First Author | Sun H | Year | 2020 |
Journal | Anal Biochem | Volume | 591 |
Pages | 113512 | PubMed ID | 31786225 |
Mgi Jnum | J:298404 | Mgi Id | MGI:6480080 |
Doi | 10.1016/j.ab.2019.113512 | Citation | Sun H, et al. (2020) Assessment of expression and specific activities of transglutaminases TG1, TG2, and FXIII-A during osteoclastogenesis. Anal Biochem 591:113512 |
abstractText | Osteoclasts are large multinucleated bone-resorbing cells derived from monocyte/macrophage lineage. Macrophage-colony stimulating factor (M-CSF) and receptor activator of nuclear factor-kappaB ligand (RANKL) drive the multi-stage osteoclastogenesis. Transglutaminases (TGs) are Ca(2+)- and thiol-dependent acyl transferases and protein crosslinking enzymes. TG enzyme family contains eight catalytically active enzymes TG1-7 and Factor XIII-A (FXIII-A). Recent studies have shown that TG1, TG2, and FXIII-A are present in osteoclasts and that TG2 and FXIII-A regulate osteoclastogenesis. In this study, we examined gene and protein expression and specific activities of TG1, TG2, and FXIII-A during osteoclastogenesis using "Hitomi peptides" in a day-by-day manner. We report that TG activities are highest in the differentiation and early fusion phases and then decrease dramatically. TG activities were upregulated by M-CSF and downregulated by addition of RANKL. FXIII-A was dramatically downregulated by RANKL, suggesting its involvement in M-CSF-mediated precursor commitment phase. TG1 and TG2 proteins were present throughout osteoclastogenesis, suggesting that they may have functions in both differentiation and fusion. In summary, the three TGs likely exert distinct functions at different stages of osteoclastogenesis. Our work also demonstrates that the "Hitomi peptides" are highly specific tools for detection of distinct TGs in a system where multiple TGs are present. |