Molecular modelling is routinely employed to assign 3D structures to collision cross sections (CCSs) derived from ion mobility mass spectrometry experiments (IM-MS). The assignment of model structures to the experimental CCSs remains an ambiguous task, where one of several methods may be used to obtain a CCS from a given set of coordinates. The most reliable of the commonly used techniques, the Trajectory Method, starts with atomic coordinates which can be accompanied by partial atomic charges, obtained using ab initio methods. Here, we use lithiated α- and β-glucose ions as exemplar molecules to detect the effect conformational modification and changes to the partial charge distribution have on computed collision cross sections. Six popular charge schemes (Mulliken, APT, CHelpG, MK, HLY and NPA) were examined in combination with three functionals (Hartree-Fock, B3LYP and M05) and five basis sets (STO-3G, 3-21G, 6-31G, 6-31+G and 6-31G ^* ) on twenty unique structures. Our findings indicate that molecular conformation makes a significant contribution to fluctuations of partial charges in Electrostatic Potential (ESP) and Mulliken charge scheme; Partial charges derived using Natural Population Analysis (NPA) and ESP methods are largely independent of functional and basis set selection; and both selection of the charge scheme and functional/basis set combination play a large role in the resultant CCS, often causing few percent fluctuations in the computed values.

In this manuscript we address an important gap in our current carbohydrate active enzyme toolbox, by developing a highly a2,6-selective (over a2,3-selective) de facto sialidase that is necessary both for glycan analysis and glycoconjugate remodeling. Both glycosidic linkages are commonly found in animal biology and each has been shown to have distinct biological function. Our approach is novel in that it harnesses the high selectivity of known glycosyltransferases ‘in reverse’ for effective hydrolysis, converting transferases to hydrolases by reaction engineering. More specifically, we demonstrate that the a2,6-specific pseudosialidase activity of Photobacterium sp. JT-ISH-224 a2,6-sialyltransferase can be used effectively for highly a2,6 selective hydrolysis on a broad range of analytes: small synthetic probes, isolated complex glycans and complex mixtures of glycoproteins.

In this manuscript we address an important gap in our current carbohydrate active enzyme toolbox, by developing a highly a2,6-selective (over a2,3-selective) de facto sialidase that is necessary both for glycan analysis and glycoconjugate remodeling. Both glycosidic linkages are commonly found in animal biology and each has been shown to have distinct biological function. Our approach is novel in that it harnesses the high selectivity of known glycosyltransferases ‘in reverse’ for effective hydrolysis, converting transferases to hydrolases by reaction engineering. More specifically, we demonstrate that the a2,6-specific pseudosialidase activity of Photobacterium sp. JT-ISH-224 a2,6-sialyltransferase can be used effectively for highly a2,6 selective hydrolysis on a broad range of analytes: small synthetic probes, isolated complex glycans and complex mixtures of glycoproteins.