Discovery of a novel caspase-1 substrate using a cell-free based protein array

2015 Fiscal Year Final Research Report

• Project/Area Number: 25290077
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: System genome science
• Research Institution: Ehime University
• Principal Investigator: SAWASAKI TATSUYA 愛媛大学, プロテオサイエンスセンター, 教授 (50314969)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: カスパーゼ１ / 無細胞 / プロテインアレイ / 基質探索 / プロテアーゼ

Outline of Final Research Achievements

We were successful to make a NC-tagged protein array consisting of about 1,000 kinds of signal transduction proteins from 300 protein kinases and 700 singe transmembrane proteins, which these proteins were fused to N-terminally FLAG tag and C-terminally biotin ligation site to detect protein cleavage by AlphaScreen technology. We screened and found a protein cleaved by capase-1 (CASP1) in the protein array. A single cleavage site in a protein was identified by alanine mutagenesis. The new protein was also cleaved in the cells that CASP1 was activated and the C-terminally cleaved fragment was located in the nucleus although the N-terminal fragment seemed to be degradated. These results suggest that this protein is a novel substrate protein for CASP1.

Free Research Field

蛋白質科学