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In recent decades, the incidence of death and morbidity due to diabetes has increased worldwide, causing a high social and economic impact. Diabetes is a major cause of blindness, kidney failure, heart attack, stroke and lower limb amputation. However, the molecular mechanisms that make the heart and kidneys the main targets of diabetes are not completely understood. To better understand the complex biochemical mechanism of diabetic cardiomyopathy, we investigated the effects of hyperglycemia with concomitant digoxin and ouabain stimulation in H9c2 cells. Total extracted proteins were analyzed by label-free LC-MS/MS, quantified by Scaffold software and validated by parallel reaction monitoring (PRM) methodology. Here, we show that the eukaryotic initiation factors (Eifs) and elongation factors (Eefs) Eif3f, Eef2 and Eif4a1 are overexpressed following cardiotonic steroid (CTS) stimulation. Similarly, the expression of four 14-3-3 proteins that play a key role in cardiac ventricular compaction was altered after CTS stimulation. In total, the expression of nine protein groups was altered in response to the stimulation of H9c2 cells. Here, the biological consequences of these changes are discussed in depth. Hyperglycemia is the main physiological condition that provokes tissue and vascular injuries in heart of diabetic patients. However, the changings at large scale in the expression of proteins of cardiomyocytes generated by this condition was not yet studied. Here we report for the first time the altered biosynthesis of nine groups of proteins of H9c2 cells activated by high glucose concentrations and by cardiotonic steroids (CTS). Furthermore, the increased biosynthesis of Eifs, Eefs and 14-3-3 protein groups by CTS, which play a crucial role in cardiomyopathies are original data reported in this work. These findings not only enhance our knowledge concerning to the effects of hyperglycemia and CTS on H9c2 cells but also indicate potential molecular targets to interfere in diabetes cardiomyopathy progression.
Erika Meneses-Romero; Lorena Hernández-Orihuela; Victoria Pando-Robles; Tomas D. López; Juan A. Oses-Prieto; Alma L. Burlingame; Cesar V.F. Batista. Quantitative proteomic analysis reveals high interference on protein expression of H9c2 cells activated with glucose and cardiotonic steroids. Journal of Proteomics 2019, 211, 103536 .
AMA StyleErika Meneses-Romero, Lorena Hernández-Orihuela, Victoria Pando-Robles, Tomas D. López, Juan A. Oses-Prieto, Alma L. Burlingame, Cesar V.F. Batista. Quantitative proteomic analysis reveals high interference on protein expression of H9c2 cells activated with glucose and cardiotonic steroids. Journal of Proteomics. 2019; 211 ():103536.
Chicago/Turabian StyleErika Meneses-Romero; Lorena Hernández-Orihuela; Victoria Pando-Robles; Tomas D. López; Juan A. Oses-Prieto; Alma L. Burlingame; Cesar V.F. Batista. 2019. "Quantitative proteomic analysis reveals high interference on protein expression of H9c2 cells activated with glucose and cardiotonic steroids." Journal of Proteomics 211, no. : 103536.
Conus snails produce venoms containing numerous peptides such as the α-conotoxins (α-CTXs), which are well-known nicotinic acetylcholine receptor (nAChR) antagonists. Thirty-eight chromatographic fractions from Conus princeps venom extract were isolated by RP-HPLC. The biological activities of 37 fractions (0.07 µg/µL) were assayed by two-electrode voltage clamp on human α7 nAChRs expressed in Xenopus laevis oocytes. Fractions F7 and F16 notably inhibited the response elicited by acetylcholine by 52.7 ± 15.2% and 59.6 ± 2.5%, respectively. Fraction F7 was purified, and an active peptide (F7-3) was isolated. Using a combination of Edman degradation, mass spectrometry, and RNASeq, we determined the sequence of peptide F7-3: AVKKTCIRSTOGSNWGRCCLTKMCHTLCCARSDCTCVYRSGKGHGCSCTS, with one hydroxyproline (O) and a free C-terminus. The average mass of this peptide, 10,735.54 Da, indicates that it is a homodimer of identical subunits, with 10 disulfide bonds in total. This peptide is clearly similar to αD-CTXs from species of the Indo-Pacific. Therefore, we called it αD-PiXXA. This toxin slowly and reversibly inhibited the ACh-induced response of the hα7 nAChR subtype, with an IC50 of 6.2 μM, and it does not affect the hα3β2 subtype at 6.5 μM.
Arisaí C. Hernández-Sámano; Andrés Falcón; Fernando Zamudio; César V.F. Batista; Jesús Emilio Michel-Morfín; Víctor Landa-Jaime; Estuardo López-Vera; Michael C. Jeziorski; Manuel B. Aguilar. αD-Conotoxins in Species of the Eastern Pacific: The Case of Conus princeps from Mexico. Toxins 2019, 11, 405 .
AMA StyleArisaí C. Hernández-Sámano, Andrés Falcón, Fernando Zamudio, César V.F. Batista, Jesús Emilio Michel-Morfín, Víctor Landa-Jaime, Estuardo López-Vera, Michael C. Jeziorski, Manuel B. Aguilar. αD-Conotoxins in Species of the Eastern Pacific: The Case of Conus princeps from Mexico. Toxins. 2019; 11 (7):405.
Chicago/Turabian StyleArisaí C. Hernández-Sámano; Andrés Falcón; Fernando Zamudio; César V.F. Batista; Jesús Emilio Michel-Morfín; Víctor Landa-Jaime; Estuardo López-Vera; Michael C. Jeziorski; Manuel B. Aguilar. 2019. "αD-Conotoxins in Species of the Eastern Pacific: The Case of Conus princeps from Mexico." Toxins 11, no. 7: 405.
Venom glands and soluble venom from the Mexican scorpion Centruroides limpidus (Karsch, 1879) were used for transcriptomic and proteomic analyses, respectively. An RNA-seq was performed by high-throughput sequencing with the Illumina platform. Approximately 80 million reads were obtained and assembled into 198,662 putative transcripts, of which 11,058 were annotated by similarity to sequences from available databases. A total of 192 venom-related sequences were identified, including Na+ and K+ channel-acting toxins, enzymes, host defense peptides, and other venom components. The most diverse transcripts were those potentially coding for ion channel-acting toxins, mainly those active on Na+ channels (NaScTx). Sequences corresponding to β- scorpion toxins active of K+ channels (KScTx) and λ-KScTx are here reported for the first time for a scorpion of the genus Centruroides. Mass fingerprint corroborated that NaScTx are the most abundant components in this venom. Liquid chromatography coupled to mass spectometry (LC-MS/MS) allowed the identification of 46 peptides matching sequences encoded in the transcriptome, confirming their expression in the venom. This study corroborates that, in the venom of toxic buthid scorpions, the more abundant and diverse components are ion channel-acting toxins, mainly NaScTx, while they lack the HDP diversity previously demonstrated for the non-buthid scorpions. The highly abundant and diverse antareases explain the pancreatitis observed after envenomation by this species.
Jimena I. Cid-Uribe; Erika P. Meneses; Cesar V. F. Batista; Ernesto Ortiz; Lourival D. Possani. Dissecting Toxicity: The Venom Gland Transcriptome and the Venom Proteome of the Highly Venomous Scorpion Centruroides limpidus (Karsch, 1879). Toxins 2019, 11, 247 .
AMA StyleJimena I. Cid-Uribe, Erika P. Meneses, Cesar V. F. Batista, Ernesto Ortiz, Lourival D. Possani. Dissecting Toxicity: The Venom Gland Transcriptome and the Venom Proteome of the Highly Venomous Scorpion Centruroides limpidus (Karsch, 1879). Toxins. 2019; 11 (5):247.
Chicago/Turabian StyleJimena I. Cid-Uribe; Erika P. Meneses; Cesar V. F. Batista; Ernesto Ortiz; Lourival D. Possani. 2019. "Dissecting Toxicity: The Venom Gland Transcriptome and the Venom Proteome of the Highly Venomous Scorpion Centruroides limpidus (Karsch, 1879)." Toxins 11, no. 5: 247.
To understand the diversity of scorpion venom, RNA from venomous glands from a sawfinger scorpion, Serradigitus gertschi, of the family Vaejovidae, was extracted and used for transcriptomic analysis. A total of 84,835 transcripts were assembled after Illumina sequencing. From those, 119 transcripts were annotated and found to putatively code for peptides or proteins that share sequence similarities with the previously reported venom components of other species. In accordance with sequence similarity, the transcripts were classified as potentially coding for 37 ion channel toxins; 17 host defense peptides; 28 enzymes, including phospholipases, hyaluronidases, metalloproteases, and serine proteases; nine protease inhibitor-like peptides; 10 peptides of the cysteine-rich secretory proteins, antigen 5, and pathogenesis-related 1 protein superfamily; seven La1-like peptides; and 11 sequences classified as “other venom components”. A mass fingerprint performed by mass spectrometry identified 204 components with molecular masses varying from 444.26 Da to 12,432.80 Da, plus several higher molecular weight proteins whose precise masses were not determined. The LC-MS/MS analysis of a tryptic digestion of the soluble venom resulted in the de novo determination of 16,840 peptide sequences, 24 of which matched sequences predicted from the translated transcriptome. The database presented here increases our general knowledge of the biodiversity of venom components from neglected non-buthid scorpions.
Maria Teresa Romero-Gutiérrez; Carlos Eduardo Santibáñez-López; Juana María Jiménez-Vargas; Cesar Vicente Ferreira Batista; Ernesto Ortiz; Lourival Domingos Possani. Transcriptomic and Proteomic Analyses Reveal the Diversity of Venom Components from the Vaejovid Scorpion Serradigitus gertschi. Toxins 2018, 10, 359 .
AMA StyleMaria Teresa Romero-Gutiérrez, Carlos Eduardo Santibáñez-López, Juana María Jiménez-Vargas, Cesar Vicente Ferreira Batista, Ernesto Ortiz, Lourival Domingos Possani. Transcriptomic and Proteomic Analyses Reveal the Diversity of Venom Components from the Vaejovid Scorpion Serradigitus gertschi. Toxins. 2018; 10 (9):359.
Chicago/Turabian StyleMaria Teresa Romero-Gutiérrez; Carlos Eduardo Santibáñez-López; Juana María Jiménez-Vargas; Cesar Vicente Ferreira Batista; Ernesto Ortiz; Lourival Domingos Possani. 2018. "Transcriptomic and Proteomic Analyses Reveal the Diversity of Venom Components from the Vaejovid Scorpion Serradigitus gertschi." Toxins 10, no. 9: 359.
The recent publication of high-throughput transcriptomic and proteomic analyses of scorpion venom glands has increased our knowledge on the biodiversity of venom components. In this contribution, we report the transcriptome of the venom gland and the proteome of the venom for the scorpion species Paravaejovis schwenkmeyeri, a member of the family Vaejovidae. We report 138 annotated transcripts encoding putative peptides/proteins with sequence identity to known venom components available from different databases. A fingerprint analysis containing the molecular masses of 212 components of the whole soluble venom revealed molecular weights of approximately 700 to 13,800 Da, with most detected proteins ranging from 1500 to 3000 Da. Amino acid sequencing of venom components by LC-MS/MS allowed the identification of fragments from 27 peptides encoded by transcripts found in the transcriptome analysis. Enzymatic assays conducted with the soluble venom fraction confirmed the presence of enzymes such as hyaluronidases and phospholipases. The database presented here increases our general knowledge on the biodiversity of venom components from neglected non-buthid scorpions.
Jimena I. Cid-Uribe; Carlos E. Santibáñez-López; Erika P. Meneses; Cesar Batista; Juana M. Jiménez-Vargas; Ernesto Ortiz; Lourival D. Possani. The diversity of venom components of the scorpion species Paravaejovis schwenkmeyeri (Scorpiones: Vaejovidae) revealed by transcriptome and proteome analyses. Toxicon 2018, 151, 47 -62.
AMA StyleJimena I. Cid-Uribe, Carlos E. Santibáñez-López, Erika P. Meneses, Cesar Batista, Juana M. Jiménez-Vargas, Ernesto Ortiz, Lourival D. Possani. The diversity of venom components of the scorpion species Paravaejovis schwenkmeyeri (Scorpiones: Vaejovidae) revealed by transcriptome and proteome analyses. Toxicon. 2018; 151 ():47-62.
Chicago/Turabian StyleJimena I. Cid-Uribe; Carlos E. Santibáñez-López; Erika P. Meneses; Cesar Batista; Juana M. Jiménez-Vargas; Ernesto Ortiz; Lourival D. Possani. 2018. "The diversity of venom components of the scorpion species Paravaejovis schwenkmeyeri (Scorpiones: Vaejovidae) revealed by transcriptome and proteome analyses." Toxicon 151, no. : 47-62.
A proteomic analysis of the soluble venom of the coral snake Micrurus pyrrhocryptus is reported in this work. The whole soluble venom was separated by RP-HPLC and the molecular weights of its components (over 100) were determined by mass spectrometry. Three main sets of components were identified, corresponding to peptides with molecular masses from 5 to 8 kDa, proteins from 12 to 16 kDa and proteins from 20 to 30 kDa. Two components were fully sequenced: one α-neurotoxic peptide of 7210 Da with slight blocking activity of the nicotinic acetylcholine receptor (nAChR) and a phospholipase A2 (PLA2) with molecular weight 13517 Da and no effect on the nAChR. PLA2 activity was evaluated for all RP-HPLC components. In addition, N-terminal sequence was obtained for eleven components using Edman degradation. Among these, three were similar to known PLA2's, six to three-finger toxins (3FTx) and one to Kunitz-type serine protease inhibitors. Two-dimensional gel electrophoresis of the venom allowed the separation of about thirty spots with components of molecular weights from 25 to 70 kDa. Seventeen spots were recovered from the gel, digested with trypsin and the corresponding peptides (85) were sequenced by MS/MS allowing identification of amino acid sequences with similarities to snake venom metalloproteases (SVMP), PLA2's, L-amino acid oxidases (LAAO), acetylcholinesterases (AChE) and serine proteases (SP). In addition, LC-MS analysis of peptides obtained from tryptic digestion of whole soluble venom allowed the identification of 695 peptides, whose amino acid sequence could correspond to at least 355 components found in other snake venoms, where C-type lectins, vespryns, zinc finger proteins, and waprins were found, among others. These results show the complexity of the venom and provide important knowledge for future work on identification and activity determination of venom components from this coral snake.
Timoteo Olamendi-Portugal; César V.F. Batista; Martha Pedraza-Escalona; Rita Restano-Cassulini; Fernando Z. Zamudio; Melisa Benard-Valle; Adolfo Rafael de Roodt; Lourival D. Possani. New insights into the proteomic characterization of the coral snake Micrurus pyrrhocryptus venom. Toxicon 2018, 153, 23 -31.
AMA StyleTimoteo Olamendi-Portugal, César V.F. Batista, Martha Pedraza-Escalona, Rita Restano-Cassulini, Fernando Z. Zamudio, Melisa Benard-Valle, Adolfo Rafael de Roodt, Lourival D. Possani. New insights into the proteomic characterization of the coral snake Micrurus pyrrhocryptus venom. Toxicon. 2018; 153 ():23-31.
Chicago/Turabian StyleTimoteo Olamendi-Portugal; César V.F. Batista; Martha Pedraza-Escalona; Rita Restano-Cassulini; Fernando Z. Zamudio; Melisa Benard-Valle; Adolfo Rafael de Roodt; Lourival D. Possani. 2018. "New insights into the proteomic characterization of the coral snake Micrurus pyrrhocryptus venom." Toxicon 153, no. : 23-31.
The soluble venom from the scorpion Tityus metuendus was characterized by various methods. In vivo experiments with mice showed that it is lethal. Extended electrophysiological recordings using seven sub-types of human voltage gated sodium channels (hNav1.1 to 1.7) showed that it contains both α- and β-scorpion toxin types. Fingerprint analysis by mass spectrometry identified over 200 distinct molecular mass components. At least 60 sub-fractions were recovered from HPLC separation. Five purified peptides were sequenced by Edman degradation, and their complete primary structures were determined. Additionally, three other peptides have had their N-terminal amino acid sequences determined by Edman degradation and reported. Mass spectrometry analysis of tryptic digestion of the soluble venom permitted the identification of the amino acid sequence of 111 different peptides. Search for similarities of the sequences found indicated that they probably are: sodium and potassium channel toxins, metalloproteinases, hyaluronidases, endothelin and angiotensin-converting enzymes, bradykinin-potentiating peptide, hypothetical proteins, allergens, other enzymes, other proteins and peptides.
Cesar Batista; J.G. Martins; R. Restano-Cassulini; F.I.V. Coronas; F.Z. Zamudio; R. Procópio; L.D. Possani. Venom characterization of the Amazonian scorpion Tityus metuendus. Toxicon 2018, 143, 51 -58.
AMA StyleCesar Batista, J.G. Martins, R. Restano-Cassulini, F.I.V. Coronas, F.Z. Zamudio, R. Procópio, L.D. Possani. Venom characterization of the Amazonian scorpion Tityus metuendus. Toxicon. 2018; 143 ():51-58.
Chicago/Turabian StyleCesar Batista; J.G. Martins; R. Restano-Cassulini; F.I.V. Coronas; F.Z. Zamudio; R. Procópio; L.D. Possani. 2018. "Venom characterization of the Amazonian scorpion Tityus metuendus." Toxicon 143, no. : 51-58.
This communication reports a further examination of venom gland transcripts and venom composition of the Mexican scorpion Thorellius atrox using RNA-seq and tandem mass spectrometry. The RNA-seq, which was performed with the Illumina protocol, yielded more than 20,000 assembled transcripts. Following a database search and annotation strategy, 160 transcripts were identified, potentially coding for venom components. A novel sequence was identified that potentially codes for a peptide with similarity to spider ω-agatoxins, which act on voltage-gated calcium channels, not known before to exist in scorpion venoms. Analogous transcripts were found in other scorpion species. They could represent members of a new scorpion toxin family, here named omegascorpins. The mass fingerprint by LC-MS identified 135 individual venom components, five of which matched with the theoretical masses of putative peptides translated from the transcriptome. The LC-MS/MS de novo sequencing allowed to reconstruct and identify 42 proteins encoded by assembled transcripts, thus validating the transcriptome analysis. Earlier studies conducted with this scorpion venom permitted the identification of only twenty putative venom components. The present work performed with more powerful and modern omic technologies demonstrates the capacity of accomplishing a deeper characterization of scorpion venom components and the identification of novel molecules with potential applications in biomedicine and the study of ion channel physiology.
Teresa Romero-Gutierrez; Esteban Peguero-Sanchez; Miguel A. Cevallos; Cesar V. F. Batista; Ernesto Ortiz; Lourival D. Possani. A Deeper Examination of Thorellius atrox Scorpion Venom Components with Omic Technologies. Toxins 2017, 9, 399 .
AMA StyleTeresa Romero-Gutierrez, Esteban Peguero-Sanchez, Miguel A. Cevallos, Cesar V. F. Batista, Ernesto Ortiz, Lourival D. Possani. A Deeper Examination of Thorellius atrox Scorpion Venom Components with Omic Technologies. Toxins. 2017; 9 (12):399.
Chicago/Turabian StyleTeresa Romero-Gutierrez; Esteban Peguero-Sanchez; Miguel A. Cevallos; Cesar V. F. Batista; Ernesto Ortiz; Lourival D. Possani. 2017. "A Deeper Examination of Thorellius atrox Scorpion Venom Components with Omic Technologies." Toxins 9, no. 12: 399.
The soluble venom from the Mexican scorpion Megacormus gertschi of the family Euscorpiidae was obtained and its biological effects were tested in several animal models. This venom is not toxic to mice at doses of 100 μg per 20 g of mouse weight, while being lethal to arthropods (insects and crustaceans), at doses of 20 μg (for crickets) and 100 μg (for shrimps) per animal. Samples of the venom were separated by high performance liquid chromatography and circa 80 distinct chromatographic fractions were obtained from which 67 components have had their molecular weights determined by mass spectrometry analysis. The N-terminal amino acid sequence of seven protein/peptides were obtained by Edman degradation and are reported. Among the high molecular weight components there are enzymes with experimentally-confirmed phospholipase activity. A pair of telsons from this scorpion species was dissected, from which total RNA was extracted and used for cDNA library construction. Massive sequencing by the Illumina protocol, followed by de novo assembly, resulted in a total of 110,528 transcripts. From those, we were able to annotate 182, which putatively code for peptides/proteins with sequence similarity to previously-reported venom components available from different protein databases. Transcripts seemingly coding for enzymes showed the richest diversity, with 52 sequences putatively coding for proteases, 20 for phospholipases, 8 for lipases and 5 for hyaluronidases. The number of different transcripts potentially coding for peptides with sequence similarity to those that affect ion channels was 19, for putative antimicrobial peptides 19, and for protease inhibitor-like peptides, 18. Transcripts seemingly coding for other venom components were identified and described. The LC/MS analysis of a trypsin-digested venom aliquot resulted in 23 matches with the translated transcriptome database, which validates the transcriptome. The proteomic and transcriptomic analyses reported here constitute the first approach to study the venom components from a scorpion species belonging to the family Euscorpiidae. The data certainly show that this venom is different from all the ones described thus far in the literature.
Carlos E. Santibáñez-López; Jimena I. Cid-Uribe; Fernando Z. Zamudio; Cesar V.F. Batista; Ernesto Ortiz; Lourival D. Possani. Venom gland transcriptomic and venom proteomic analyses of the scorpion Megacormus gertschi Díaz-Najera, 1966 (Scorpiones: Euscorpiidae: Megacorminae). Toxicon 2017, 133, 95 -109.
AMA StyleCarlos E. Santibáñez-López, Jimena I. Cid-Uribe, Fernando Z. Zamudio, Cesar V.F. Batista, Ernesto Ortiz, Lourival D. Possani. Venom gland transcriptomic and venom proteomic analyses of the scorpion Megacormus gertschi Díaz-Najera, 1966 (Scorpiones: Euscorpiidae: Megacorminae). Toxicon. 2017; 133 ():95-109.
Chicago/Turabian StyleCarlos E. Santibáñez-López; Jimena I. Cid-Uribe; Fernando Z. Zamudio; Cesar V.F. Batista; Ernesto Ortiz; Lourival D. Possani. 2017. "Venom gland transcriptomic and venom proteomic analyses of the scorpion Megacormus gertschi Díaz-Najera, 1966 (Scorpiones: Euscorpiidae: Megacorminae)." Toxicon 133, no. : 95-109.
Aedes-borne viruses are responsible for high-impact neglected tropical diseases and unpredictable outbreaks such as the ongoing Zika epidemics. Aedes mosquitoes spread different arboviruses such as Dengue virus (DENV), Chikungunya virus (CHIKV), and Zika virus, among others, and are responsible for the continuous emergence and reemergence of these pathogens. These viruses have complex transmission cycles that include two hosts, namely the Aedes mosquito as a vector and susceptible vertebrate hosts. Human infection with arboviruses causes diseases that range from subclinical or mild to febrile diseases, encephalitis, and hemorrhagic fever. Infected mosquitoes do not show detectable signs of disease, even though the virus maintains a lifelong persistent infection. The infection of the Aedes mosquito by viruses involves a molecular crosstalk between cell and viral proteins. An understanding of how mosquito vectors and viruses interact is of fundamental interest, and it also offers novel perspectives for disease control. In recent years, mass spectrometry (MS)-based strategies in combination with bioinformatics have been successfully applied to identify and quantify global changes in cellular proteins, lipids, peptides, and metabolites in response to viral infection. Although the information about proteomics in the Aedes mosquito is limited, the information that has been reported can set up the basis for future studies. This review reflects how MS-based approaches have extended our understanding of Aedes mosquito biology and the development of DENV and CHIKV infection in the vector. Finally, this review discusses future challenges in the field.
Victoria Pando-Robles; Cesar V. Batista. Aedes-Borne Virus–Mosquito Interactions: Mass Spectrometry Strategies and Findings. Vector-Borne and Zoonotic Diseases 2017, 17, 361 -375.
AMA StyleVictoria Pando-Robles, Cesar V. Batista. Aedes-Borne Virus–Mosquito Interactions: Mass Spectrometry Strategies and Findings. Vector-Borne and Zoonotic Diseases. 2017; 17 (6):361-375.
Chicago/Turabian StyleVictoria Pando-Robles; Cesar V. Batista. 2017. "Aedes-Borne Virus–Mosquito Interactions: Mass Spectrometry Strategies and Findings." Vector-Borne and Zoonotic Diseases 17, no. 6: 361-375.
Venom from male and female scorpions of the species Centruroides limpidus were separated by HPLC and their molecular masses determined by mass spectrometry. The relative concentration of components eluting in equivalent retention times from the HPLC column shows some differences. A new peptide with 29 amino acids, cross-linked by three disulfide bonds was found in male scorpions and its structure determined. Another unknown peptide present in female venom, with sequence identity similar to K+-channel blocking peptide was isolated. This peptide contains 39 amino acid residues linked by three disulfide bonds. Due to sequence similarities, a systematic number (αKTx2.18) was assigned. Venom from male and female scorpions was separated by Sephadex G-50 gel filtration. Components of fraction I of this chromatogram were analyzed by two-dimensional gel electrophoresis and 41 spots were selected (20 from female and 21 from male). The spots were excised from the gel, enzymatically digested and sequenced by LC-MS/MS. This procedure allowed the identification of several proteins containing similar amino acid sequence of other known proteins registered on UniProt database. Among these proteins the presence of metalloproteinases (proteolytic enzymes), hyaluronidases and phosphatases were experimentally determined and shown to be present in both venom samples. The results shown here should help further work aimed at fully identification of the structure and function of venom components form C. limpidus male and female scorpions.
Jimena Isaias Cid Uribe; Juana Maria Jiménez Vargas; Cesar Vicente Ferreira Batista; Fernando Zamudio Zuñiga; Lourival Domingos Possani. Comparative proteomic analysis of female and male venoms from the Mexican scorpion Centruroides limpidus: Novel components found. Toxicon 2017, 125, 91 -98.
AMA StyleJimena Isaias Cid Uribe, Juana Maria Jiménez Vargas, Cesar Vicente Ferreira Batista, Fernando Zamudio Zuñiga, Lourival Domingos Possani. Comparative proteomic analysis of female and male venoms from the Mexican scorpion Centruroides limpidus: Novel components found. Toxicon. 2017; 125 ():91-98.
Chicago/Turabian StyleJimena Isaias Cid Uribe; Juana Maria Jiménez Vargas; Cesar Vicente Ferreira Batista; Fernando Zamudio Zuñiga; Lourival Domingos Possani. 2017. "Comparative proteomic analysis of female and male venoms from the Mexican scorpion Centruroides limpidus: Novel components found." Toxicon 125, no. : 91-98.
Venom gland transcriptomic and proteomic analyses have improved our knowledge on the diversity of the heterogeneous components present in scorpion venoms. However, most of these studies have focused on species from the family Buthidae. To gain insights into the molecular diversity of the venom components of scorpions belonging to the family Superstitioniidae, one of the neglected scorpion families, we performed a transcriptomic and proteomic analyses for the species Superstitionia donensis. The total mRNA extracted from the venom glands of two specimens was subjected to massive sequencing by the Illumina protocol, and a total of 219,073 transcripts were generated. We annotated 135 transcripts putatively coding for peptides with identity to known venom components available from different protein databases. Fresh venom collected by electrostimulation was analyzed by LC-MS/MS allowing the identification of 26 distinct components with sequences matching counterparts from the transcriptomic analysis. In addition, the phylogenetic affinities of the found putative calcins, scorpines, La1-like peptides and potassium channel κ toxins were analyzed. The first three components are often reported as ubiquitous in the venom of different families of scorpions. Our results suggest that, at least calcins and scorpines, could be used as molecular markers in phylogenetic studies of scorpion venoms.
Carlos E. Santibáñez-López; Jimena I. Cid-Uribe; Cesar V. F. Batista; Ernesto Ortiz; Lourival D. Possani. Venom Gland Transcriptomic and Proteomic Analyses of the Enigmatic Scorpion Superstitionia donensis (Scorpiones: Superstitioniidae), with Insights on the Evolution of Its Venom Components. Toxins 2016, 8, 367 .
AMA StyleCarlos E. Santibáñez-López, Jimena I. Cid-Uribe, Cesar V. F. Batista, Ernesto Ortiz, Lourival D. Possani. Venom Gland Transcriptomic and Proteomic Analyses of the Enigmatic Scorpion Superstitionia donensis (Scorpiones: Superstitioniidae), with Insights on the Evolution of Its Venom Components. Toxins. 2016; 8 (12):367.
Chicago/Turabian StyleCarlos E. Santibáñez-López; Jimena I. Cid-Uribe; Cesar V. F. Batista; Ernesto Ortiz; Lourival D. Possani. 2016. "Venom Gland Transcriptomic and Proteomic Analyses of the Enigmatic Scorpion Superstitionia donensis (Scorpiones: Superstitioniidae), with Insights on the Evolution of Its Venom Components." Toxins 8, no. 12: 367.
Disulfide C-terminal loop fragments derived from AMPs and the presence of peptidases have been previously reported in the skin secretions of different amphibians. However, there are only a few studies on the identification of enzymes in frog skin secretion based on the primary structure of these proteins. Similarly, relative little data exists regarding the identification of disulfide C-terminal loops at large scale. Therefore, a comprehensive study on this issue certainly could bring in much more information for understanding this molecular process and its biochemical consequences. Thus, the aim of this work was to characterize the presence of disulfide C-terminal loop fragments of AMPs and identify the proteins and probable enzymes present in the completely unknown secretion contents of the frog Lithobates spectabilis. For this purpose, high-resolution mass spectrometry was applied to analyze the skin secretions processed by two different protocols: (1) using a cocktail of enzymatic inhibitors and 2) without any protease inhibitors, maintaining the solution for 2 hours at 10°C. Results from procedure-1, revealed 122 molecular masses, whereas procedure-2 permitted 253 different molecular masses to be identified. Fifty-nine peptides including 22 disulfide C-terminal loop-containing peptides were obtained following procedure-2. Polyacrylamide gel electrophoresis separation, tryptic digestion and LC-MS/MS were used for "de novo" sequencing of 111 different peptides and the unequivocal identification of fifteen proteins including at least three different peptidases. Additionally, it was possible to fully sequence eight peptides, including a ranatuerin-related peptide identified here as Spectabilin, that was subsequently chemically synthesized and showed high antibacterial, antiparasitic and cytotoxic activities.
Griselda Demesa Balderrama; Erika Patricia Meneses; Lorena Hernández Orihuela; Victoria Pando- Robles; María Carmen Rodríguez; Carolina Barrientos-Salcedo; Manuel B. Aguilar; Cesar V. F. Batista. A comprehensive proteomic study of the skin secretions of the frog Lithobates spectabilis. Protein & Peptide Letters 2016, 23, 597 -611.
AMA StyleGriselda Demesa Balderrama, Erika Patricia Meneses, Lorena Hernández Orihuela, Victoria Pando- Robles, María Carmen Rodríguez, Carolina Barrientos-Salcedo, Manuel B. Aguilar, Cesar V. F. Batista. A comprehensive proteomic study of the skin secretions of the frog Lithobates spectabilis. Protein & Peptide Letters. 2016; 23 (7):597-611.
Chicago/Turabian StyleGriselda Demesa Balderrama; Erika Patricia Meneses; Lorena Hernández Orihuela; Victoria Pando- Robles; María Carmen Rodríguez; Carolina Barrientos-Salcedo; Manuel B. Aguilar; Cesar V. F. Batista. 2016. "A comprehensive proteomic study of the skin secretions of the frog Lithobates spectabilis." Protein & Peptide Letters 23, no. 7: 597-611.
A complete mass spectrometry analysis of venom components from male and female scorpions of the species Rhophalurus junceus of Cuba is reported. In the order of 200 individual molecular masses were identified in both venoms, from which 63 are identical in male and females genders. It means that a significant difference of venom components exists between individuals of different sexes, but the most abundant components are present in both sexes. The relative abundance of identical components is different among the genders. Three well defined groups of different peptides were separated and identified. The first group corresponds to peptides with molecular masses of 1000-2000 Da; the second to peptides with 3500-4500 Da molecular weight, and the third with 6500-8000 Da molecular weights. A total of 86 peptides rich in disulfide bridges were found in the venoms, 27 with three disulfide bridges and 59 with four disulfide bridges. LC-MS/MS analysis allowed the identification and amino acid sequence determination of 31 novel peptides in male venom. Two new putative K(+)-channel peptides were sequences by Edman degradation. They contain 37 amino acid residues, packed by three disulfide bridges and were assigned the systematic numbers: α-KTx 1.18 and α-KTx 2.15.
Rodolfo Rodríguez-Ravelo; Cesar V.F. Batista; Fredy I.V. Coronas; Fernando Z. Zamudio; Lorena Hernández-Orihuela; Georgina Espinosa-López; Ariel Ruiz-Urquiola; Lourival D. Possani. Comparative proteomic analysis of male and female venoms from the Cuban scorpion Rhopalurus junceus. Toxicon 2015, 107, 327 -334.
AMA StyleRodolfo Rodríguez-Ravelo, Cesar V.F. Batista, Fredy I.V. Coronas, Fernando Z. Zamudio, Lorena Hernández-Orihuela, Georgina Espinosa-López, Ariel Ruiz-Urquiola, Lourival D. Possani. Comparative proteomic analysis of male and female venoms from the Cuban scorpion Rhopalurus junceus. Toxicon. 2015; 107 ():327-334.
Chicago/Turabian StyleRodolfo Rodríguez-Ravelo; Cesar V.F. Batista; Fredy I.V. Coronas; Fernando Z. Zamudio; Lorena Hernández-Orihuela; Georgina Espinosa-López; Ariel Ruiz-Urquiola; Lourival D. Possani. 2015. "Comparative proteomic analysis of male and female venoms from the Cuban scorpion Rhopalurus junceus." Toxicon 107, no. : 327-334.
Although the primary physiological effects produced by scorpion toxins are already well known, most of the secondary molecular events following scorpion neurotoxins-ion channel interactions are poorly understood and described. For this reason, we used a proteomic approach to determine the changes in relative protein abundance in F11 mouse neuroblastoma cells treated with Cn2, the major β-toxin from the venom of the scorpion Centruroides noxius Hoffmann. Here we show that the relative abundance of 24 proteins changed after Cn2 treatment. Proteins related to protection from apoptosis and cell survival, as well as those involved in cell morphology and some translation elongation factors were diminished. By contrast, proteins associated with energy metabolism were increased. Additionally, results of western immunoblots confirmed the preference of action towards some special targets. These results suggest that Cn2 could modify the neuronal structure and induce apoptosis and reduction of the proliferation and cell survival. To support this conclusion we directly measured the Cn2 effect on cell proliferation, division and apoptosis. Our results open new avenues for continuing the studies aimed at better understanding the envenomation process caused by scorpion stings. The purpose of this work was to identify which proteins, apart from the ion-channels, are involved in the envenomation process in order to develop possible strategies to circumvent the deleterious effects caused by the toxic peptides of the venom. For this reason, we characterized the early changes in the proteome of F11 cells induced by Cn2, the major toxin of the New World scorpion C. noxius Hoffmann, using 2D-PAGE and LC-MS/MS. We identified 24 proteins which relative abundance is modified after the Cn2 treatment. Among these, proteins related with apoptosis protection, cell survival, neuronal morphology and some translation elongation factors were diminished, whereas proteins associated with energy metabolism were increased.
Martha Pedraza Escalona; Cesar V.F. Batista; Rita Restano Cassulini; Marisol Sandoval Rios; Fredy I. Coronas; Lourival D. Possani. A proteomic analysis of the early secondary molecular effects caused by Cn2 scorpion toxin on neuroblastoma cells. Journal of Proteomics 2014, 111, 212 -223.
AMA StyleMartha Pedraza Escalona, Cesar V.F. Batista, Rita Restano Cassulini, Marisol Sandoval Rios, Fredy I. Coronas, Lourival D. Possani. A proteomic analysis of the early secondary molecular effects caused by Cn2 scorpion toxin on neuroblastoma cells. Journal of Proteomics. 2014; 111 ():212-223.
Chicago/Turabian StyleMartha Pedraza Escalona; Cesar V.F. Batista; Rita Restano Cassulini; Marisol Sandoval Rios; Fredy I. Coronas; Lourival D. Possani. 2014. "A proteomic analysis of the early secondary molecular effects caused by Cn2 scorpion toxin on neuroblastoma cells." Journal of Proteomics 111, no. : 212-223.
Dengue is an important and growing public health problem worldwide with an estimated 100million new clinical cases annually. Currently, no licensed drug or vaccine is available. During natural infection in humans, liver cells constitute one of the main targets of dengue virus (DENV) replication. However, a clear understanding of dengue pathogenesis remains elusive. In order to gain a better reading of the cross talk between virus and host cell proteins, we used a proteomics approach to analyze the host response to DENV infection in a hepatic cell line Huh-7. Differences in proteome expression were assayed 24h post-infection using label-free LC–MS. Quantitative analysis revealed 155 differentially expressed proteins, 64 of which were up-regulated and 91 down-regulated. These results reveal an important decrease in the expression of enzymes involved in the glycolytic pathway, citrate cycle, and pyruvate metabolism. This study provides large-scale quantitative information regarding protein expression in the early stages of infection that should be useful for better compression of the pathogenesis of dengue.Biological significanceDengue infection involves alterations in the homeostasis of the host cell. Defining the interactions between virus and cell proteins should provide a better understanding of how viruses propagate and cause disease. Here, we present for the first time the proteomic analysis of hepatocytes (Huh-7 cells) infected with DENV-2 by label-free LC–MS.This article is part of a Special Issue entitled: Proteomics, mass spectrometry and peptidomics, Cancun 2013. Guest Editors: César López-Camarillo, Victoria Pando-Robles and Bronwyn Jane Barkla
Victoria Pando-Robles; Juan A. Oses-Prieto; Myriam Rodríguez-Gandarilla; Erika Meneses-Romero; Alma L. Burlingame; Cesar V.F. Batista. Quantitative proteomic analysis of Huh-7 cells infected with Dengue virus by label-free LC–MS. Journal of Proteomics 2014, 111, 16 -29.
AMA StyleVictoria Pando-Robles, Juan A. Oses-Prieto, Myriam Rodríguez-Gandarilla, Erika Meneses-Romero, Alma L. Burlingame, Cesar V.F. Batista. Quantitative proteomic analysis of Huh-7 cells infected with Dengue virus by label-free LC–MS. Journal of Proteomics. 2014; 111 ():16-29.
Chicago/Turabian StyleVictoria Pando-Robles; Juan A. Oses-Prieto; Myriam Rodríguez-Gandarilla; Erika Meneses-Romero; Alma L. Burlingame; Cesar V.F. Batista. 2014. "Quantitative proteomic analysis of Huh-7 cells infected with Dengue virus by label-free LC–MS." Journal of Proteomics 111, no. : 16-29.
This communication reports the results of proteomic, transcriptomic, biochemical and electrophysiological analysis of the soluble venom and venom glands of the Mexican centipede Scolopendra viridis Say (here thereafter abbreviated S. viridis). Separation of the soluble venom permitted to obtain 54 different fractions, from which a mass finger printing analysis permitted the identification of at least 86 components, where 70% of the molecules have low molecular masses. Two-dimensional electrophoretic separation of this venom revealed the presence of about forty proteins with molecular weights ranging from 17 to 58kDa. The novo sequencing of 149 peptides obtained by LC-MS/MS from the 2D-gels showed the presence of proteins with amino acid sequences similar to several enzymes and venom allergens type 3. Furthermore, a total of 180 sequences were obtained from a cDNA library prepared with two venomous glands. From this, 155 sequences correspond to complete genes containing more than 200 base pairs each. Comparative sequence analyses of these sequences indicated the presence of different types of enzymes and toxin-like genes. Two proteins with molecular weights around 37,000 and 42,000Da were shown to contain hyaluronidase activity. Electrophysiological assays performed with soluble venom show that it decreases mammalian sodium channel currents. Animal venoms of Scolopendra species have been scarcely studied, although they have been reported to contain several bioactive compounds, some of which with potential therapeutic interest. The Mexican centipede S. viridis contains a powerful venom, capable of inflicting immediate effects on their preys. This communication is focused on the identification and description of a proteomic and transcriptomic analysis of the protein components of this venom. Several amino acid sequences similar to reported enzymes are the principal components in the S. viridis venom, but also a low number of toxins were identified. This knowledge should contribute to the understanding of the pharmacological effects caused by bites of this centipede species.
Lidia González-Morales; Martha Pedraza-Escalona; Elia Diego-Garcia; Rita Restano-Cassulini; Cesar V.F. Batista; Maria Del Carmen Gutiérrez; Lourival D. Possani. Proteomic characterization of the venom and transcriptomic analysis of the venomous gland from the Mexican centipede Scolopendra viridis. Journal of Proteomics 2014, 111, 224 -237.
AMA StyleLidia González-Morales, Martha Pedraza-Escalona, Elia Diego-Garcia, Rita Restano-Cassulini, Cesar V.F. Batista, Maria Del Carmen Gutiérrez, Lourival D. Possani. Proteomic characterization of the venom and transcriptomic analysis of the venomous gland from the Mexican centipede Scolopendra viridis. Journal of Proteomics. 2014; 111 ():224-237.
Chicago/Turabian StyleLidia González-Morales; Martha Pedraza-Escalona; Elia Diego-Garcia; Rita Restano-Cassulini; Cesar V.F. Batista; Maria Del Carmen Gutiérrez; Lourival D. Possani. 2014. "Proteomic characterization of the venom and transcriptomic analysis of the venomous gland from the Mexican centipede Scolopendra viridis." Journal of Proteomics 111, no. : 224-237.
Recombinant wild-pyrazinamidase from H37Rv Mycobacterium tuberculosis was analyzed by gel electrophoresis under differential reducing conditions to evaluate its quaternary structure. PZAse was fractionated by size exclusion chromatography under non-reducing conditions. PZAse activity was measured and mass spectrometry analysis was performed to determine the identity of proteins by de novo sequencing and to determine the presence of disulfide bonds. This study confirmed that M. tuberculosis wild type PZAse was able to form homo-dimers in vitro. Homo-dimers showed a slightly lower specific PZAse activity compared to monomeric PZAse. PZAse dimers were dissociated into monomers in response to reducing conditions. Mass spectrometry analysis confirmed the existence of disulfide bonds (C72–C138 and C138–C138) stabilizing the quaternary structure of the PZAse homo-dimer.
Daniel Rueda; Patricia Sheen; Robert H. Gilman; Carlos Bueno; Marco Santos; Victoria Pando-Robles; Cesar V. Batista; Mirko Zimic. Nicotinamidase/pyrazinamidase of Mycobacterium tuberculosis forms homo-dimers stabilized by disulfide bonds. Tuberculosis 2014, 94, 644 -648.
AMA StyleDaniel Rueda, Patricia Sheen, Robert H. Gilman, Carlos Bueno, Marco Santos, Victoria Pando-Robles, Cesar V. Batista, Mirko Zimic. Nicotinamidase/pyrazinamidase of Mycobacterium tuberculosis forms homo-dimers stabilized by disulfide bonds. Tuberculosis. 2014; 94 (6):644-648.
Chicago/Turabian StyleDaniel Rueda; Patricia Sheen; Robert H. Gilman; Carlos Bueno; Marco Santos; Victoria Pando-Robles; Cesar V. Batista; Mirko Zimic. 2014. "Nicotinamidase/pyrazinamidase of Mycobacterium tuberculosis forms homo-dimers stabilized by disulfide bonds." Tuberculosis 94, no. 6: 644-648.
Here we show for the first time that the venom from an elapid (Micrurus fulvius) contains three finger toxin (3FTxs) peptides with low toxicity but high content of lethal phospholipases A2 (PLA2). The intravenous venom LD50 in mice was 0.3μg/g. Fractionation on a C18 column yielded 22 fractions; in terms of abundance, 58.3% of them were components of 13-14kDa and 24.9% were molecules of 6-7kDa. Two fractions with PLA2 activity represented 33.4% of the whole venom and were the most lethal fractions. Fractions with low molecular mass (<7000Da) partially and reversibly blocked the nicotinic acetylcholine receptor (nAChR), with the exception of one that blocked it completely. The fraction that blocked 100% contained two protein species whose dose-response was determined; the IC50s were 13±1 and 9.5±0.3nM. Despite the apparent effect on nAChR none of the low molecular mass fractions were lethal in mice, at concentrations of 1μg/g. From 2D-PAGE and LC-MS/MS, we identified fourteen species of PLA2, four protein species of C-type lectin, three zinc metalloproteinases, one phosphodiesterase and one 3FTx. The N-terminal amino acid sequence of fractions with biological interest was obtained.
Irene Vergara; Martha Pedraza-Escalona; Dayanira Paniagua; Rita Restano-Cassulini; Fernando Zamudio; Cesar V.F. Batista; Lourival D. Possani; Alejandro Alagón. Eastern coral snake Micrurus fulvius venom toxicity in mice is mainly determined by neurotoxic phospholipases A2. Journal of Proteomics 2014, 105, 295 -306.
AMA StyleIrene Vergara, Martha Pedraza-Escalona, Dayanira Paniagua, Rita Restano-Cassulini, Fernando Zamudio, Cesar V.F. Batista, Lourival D. Possani, Alejandro Alagón. Eastern coral snake Micrurus fulvius venom toxicity in mice is mainly determined by neurotoxic phospholipases A2. Journal of Proteomics. 2014; 105 ():295-306.
Chicago/Turabian StyleIrene Vergara; Martha Pedraza-Escalona; Dayanira Paniagua; Rita Restano-Cassulini; Fernando Zamudio; Cesar V.F. Batista; Lourival D. Possani; Alejandro Alagón. 2014. "Eastern coral snake Micrurus fulvius venom toxicity in mice is mainly determined by neurotoxic phospholipases A2." Journal of Proteomics 105, no. : 295-306.
Combined quantum mechanical and molecular mechanical (QM/MM) calculations were used to explore the electron pathway involved in the suicide inactivation of cytochrome P450BM3 from Bacillus megaterium. The suicide inactivation is a common phenomenon observed for heme peroxidases, in which the enzyme is inactivated as a result of self-oxidation mediated by highly oxidizing enzyme intermediates formed during the catalytic cycle. The selected model was a mutant comprising only the heme domain (CYPBM3 21B3) that had been previously evolved to efficiently catalyze hydroxylation reactions with hydrogen peroxide (H2O2) as electron acceptor. An extensive mapping of residues involved in electron transfer routes was obtained from density functional calculations on activated heme (i.e. Compound I) and selected amino acid residues. Identification of oxidizable residues (electron donors) was performed by selectively activating/deactivating different quantum regions. This method allowed a rational identification of key oxidizable targets in order to replace them for less oxidizable residues by site-directed mutagenesis. The residues W96 and F405 were consistently predicted by the QM/MM electron pathway to hold high spin density; single and double mutants of P450BM3 on these positions (W96A, F405L, W96A/F405L) resulted in a more stable variants in the presence of hydrogen peroxide, displaying a similar reaction rate than P450BM3 21B3. Furthermore, mass spectrometry confirmed these oxidation sites and corroborated the possible routes described by QM/MM electron transfer (ET) pathways.
Abraham Vidal-Limon; Sergio A. Aguila; Marcela Ayala; Cesar V. Batista; Rafael Vazquez-Duhalt. Peroxidase activity stabilization of cytochrome P450BM3 by rational analysis of intramolecular electron transfer. Journal of Inorganic Biochemistry 2013, 122, 18 -26.
AMA StyleAbraham Vidal-Limon, Sergio A. Aguila, Marcela Ayala, Cesar V. Batista, Rafael Vazquez-Duhalt. Peroxidase activity stabilization of cytochrome P450BM3 by rational analysis of intramolecular electron transfer. Journal of Inorganic Biochemistry. 2013; 122 ():18-26.
Chicago/Turabian StyleAbraham Vidal-Limon; Sergio A. Aguila; Marcela Ayala; Cesar V. Batista; Rafael Vazquez-Duhalt. 2013. "Peroxidase activity stabilization of cytochrome P450BM3 by rational analysis of intramolecular electron transfer." Journal of Inorganic Biochemistry 122, no. : 18-26.