Cyclin M (CNNM1-4) proteins maintain cellular and body magnesium (Mg²⁺) homeostasis. Using various biochemical approaches, we have identified members of the CNNM family as direct interacting partners of ADP-ribosylation factor-like GTPase 15 (ARL15), a small GTP-binding protein. ARL15 interacts with CNNMs at their carboxyl-terminal conserved cystathionine-β-synthase (CBS) domains. In silico modeling of the interaction between CNNM2 and ARL15 supports that the small GTPase specifically binds the CBS1 and CNBH domains. Immunocytochemical experiments demonstrate that CNNM2 and ARL15 co-localize in the kidney, with both proteins showing subcellular localization in the endoplasmic reticulum, Golgi apparatus and the plasma membrane. Most importantly, we found that ARL15 is required for forming complex N-glycosylation of CNNMs. Overexpression of ARL15 promotes complex N-glycosylation of CNNM3. Mg²⁺ uptake experiments with a stable isotope demonstrate that there is a significant increase of ²⁵Mg²⁺ uptake upon knockdown of ARL15 in multiple kidney cancer cell lines. Altogether, our results establish ARL15 as a novel negative regulator of Mg²⁺ transport by promoting the complex N-glycosylation of CNNMs.

Although Ag nanoparticles (NPs) have been widely applied in daily life and in biomedical and industrial fields, there is a demand for Ag-based bimetallic nanoalloys (NAs), such as AgCu and AgFe, due to their enhanced antibacterial efficacy and reduced Ag consumption. In this work, we present a comparison study on the antibacterial efficacy and cytotoxicity rates of Ag NPs and AgCu and AgFe NAs to L929 mouse fibroblast cells using the CCK-8 technique based on the relative cell viability. The concept of the minimum death concentration (MDC) is introduced to estimate the cytotoxicity to the cells. It is found that the minimum inhibitory concentrations (MICs) of the NPs against E. coli and S. aureus decrease with the addition of both Cu and Fe. There is a strong correlation between the MDC and MIC, implying that the mechanisms of both antibacterial efficacy and cytotoxicity are similar. The enhanced antibacterial efficacy to bacteria and cytotoxicity toward the cell are attributed to Ag⁺ release. The following order is found for both the MIC and MDC: AgFe < AgCu < Ag NPs. However, there is no cytotoxicity to the L929 cells for AgFe and AgCu NAs at their MIC Ag concentrations against S. aureus.

Cyclin M (CNNM1-4) proteins maintain cellular and body magnesium (Mg ²⁺ ) homeostasis. Using various biochemical approaches, we have identified members of the CNNM family as direct interacting partners of ADP-ribosylation factor-like protein 15 (ARL15), a small GTP-binding protein. ARL15 interacts with CNNMs at their carboxyl-terminal conserved cystathionine-β-synthase (CBS) domains. In silico modeling of the interaction using the reported structures of both CNNM2 and ARL15 supports that the small GTPase specifically binds the CBS1 domain. Immunocytochemical experiments demonstrate that CNNM2 and ARL15 co-localize in the kidney, with both proteins showing subcellular localization in the Golgi-apparatus. Most importantly, we found that ARL15 is required for forming complex N-glycosylation of CNNMs. Overexpression of ARL15 promotes complex N-glycosylation of CNNM3. Mg ²⁺ uptake experiments with a stable isotope demonstrate that there is a significant increase of ²⁵ Mg ²⁺ uptake upon knockdown of ARL15 in multiple kidney cancer cell lines. Altogether, our results establish ARL15 as a novel negative regulator of Mg ²⁺ transport by promoting the complex N-glycosylation of CNNMs.