التفاصيل البيبلوغرافية
| العنوان: |
Abstract 267: Identification of Proteolytic Substrates for ADAMTS7 Using Terminal Amine Isotope Labeling of Substrates |
| المؤلفون: |
Zhang, Xuan, Klein, Theo, Cui, Jian, Ou, Kristy, Hinkle, Christine, Li, Wenjun, Overall, Christopher M, Reilly, Muredach P |
| المصدر: |
Arteriosclerosis, Thrombosis, and Vascular Biology ; volume 36, issue suppl_1 ; ISSN 1079-5642 1524-4636 |
| بيانات النشر: |
Ovid Technologies (Wolters Kluwer Health) |
| سنة النشر: |
2016 |
| الوصف: |
ADAMTS7 , a member of the disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) family, has been recently identified as a novel genetic locus for coronary artery disease. Although a handful of ADAMTS7 substrates have been identified through traditional yeast two-hybrid screening and standard proteomic profiling, the molecular mechanisms of ADAMTS7 contribution to atherosclerosis have not been fully explored in the context of its proteolytic action. Here we utilized terminal amine isotope labeling of substrates (TAILS), a cutting-edge proteomic approach to enrich N-terminal peptides, enable systemic characterization of proteolytic events and substrate identification for ADAMTS7. Briefly, primary vascular smooth muscle cells (SMC) from ADAMTS7 WT and KO mice were treated with TNFα to stimulated endogenous ADAMTS7 expression, followed by TAILS using dimethyl labeling in cellular proteome and secretome. In 3 biological replicates, we identified 1,720 and 1,616 quantified N-terminal peptides in SMC proteome and secretome, which composed of 415 and 122 unique N-termini of mature protein as well as 1,185 and 1,161 cleaved neo-N-termini driven by proteolytic processing. As expected, most of acetylated peptides in SMC proteome and secretome are natural N-termini of mature proteins (82% and 77%, respectively) while free N-terminal peptides are remarkably enriched with cleaved neo-N termini (85% and 96%, respectively). Importantly, 106 and 86 neo-N-termini (corresponding to 65 and 54 unique proteins) were enriched in ADAMTS7 WT SMC proteome and secretome respectively (WT/KO isotope ratio >2), suggesting that these proteins underwent elevated proteolysis in ADAMTS WT SMCs and thus are proteolytic substrate candidates for ADAMTS7. Interestingly, gene pathway analysis indicates that these proteins are enriched with genes involved in proteinaceous extracellular matrix and extracellular region, such as vimentin, fibulin and MMPs. Biochemistry studies are ongoing to validate these ADAMTS7 substrate candidates. ... |
| نوع الوثيقة: |
article in journal/newspaper |
| اللغة: |
English |
| DOI: |
10.1161/atvb.36.suppl_1.267 |
| الاتاحة: |
http://dx.doi.org/10.1161/atvb.36.suppl_1.267 |
| رقم الانضمام: |
edsbas.4821565C |
| قاعدة البيانات: |
BASE |