Detection and identification of potential transglutaminase 2 substrates in the mouse renal glomeruli
| Autor: | Yukio Yuzawa, Kazuo Takahashi, Hisateru Yamaguchi, Yoshimasa Ito, Kiyotaka Hitomi, Hideki Tatsukawa |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Tissue transglutaminase Kidney Glomerulus Biophysics Peptide Glomerulus (kidney) urologic and male genital diseases Biochemistry Gene Expression Regulation Enzymologic Nephropathy Gene Knockout Techniques Mice 03 medical and health sciences GTP-Binding Proteins medicine Animals Protein Glutamine gamma Glutamyltransferase 2 Molecular Biology chemistry.chemical_classification Transglutaminases 030102 biochemistry & molecular biology biology urogenital system Chemistry Glomerulonephritis medicine.disease Mice Inbred C57BL 030104 developmental biology Enzyme medicine.anatomical_structure Knockout mouse biology.protein Peptides Protein Binding Avidin |
| Zdroj: | Archives of Biochemistry and Biophysics. 660:11-19 |
| ISSN: | 0003-9861 |
| DOI: | 10.1016/j.abb.2018.10.001 |
| Popis: | The glomerulus primarily comprises mesangial cells, glomerular microvascular endothelial cells, and podocytes. IgA nephropathy is the most common primary glomerulonephritis worldwide and has a risk of progression to end-stage renal disease. IgA nephropathy is characterized by predominant IgA deposition in the glomerular mesangial area, where TG2 is significantly enhanced. Therefore, identification of glomerular TG2 substrates is the first step in elucidating the role of TG2 as a crosslinking enzyme during disease progression. To clarify potential glomerular TG2 substrates, and to establish a procedure for substrate identification, we attempted to identify those molecules using normal mouse glomeruli. Extracts from mouse glomerular and non-glomerular fractions were treated with our established biotin-labeled substrate peptide, which specifically crosslinks to the lysine-donor substrates depending on TG2 activity. Peptide-incorporated proteins were then purified using avidin resin and identified via mass spectrometry. In parallel, we performed the identification using corresponding samples from TG2 knockout mice. Consequently, potential TG2 substrates were separately identified in glomerular and non-glomerular fractions. They were mainly identified as novel TG2 substrates and partly include the well-known substrates. These results potentially provide novel insights into the mechanism underlying IgA nephropathy and may help elucidate the physiological functions of TG2. |
| Databáze: | OpenAIRE |
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