Research Project
Grant-in-Aid for Challenging Exploratory Research
Parkin, a gene product mutated in familial Parkinsonism, is essential for elimination of damaged mitochondria. Recent progress has revealed that phosphorylation of both Parkin and ubiquitin by PINK1 are crucial for their function. However, the mechanism by which phosphorylated ubiquitin associates with and activates Parkin remains largely unknown.Here, we analyze interactions between phosphorylated forms of both Parkin and ubiquitin at a spatial resolution of the amino acid residue by site-specific photo-crosslinking. We reveal that IBR domain along with RING1 domain of Parkin preferentially binds to ubiquitin in a phosphorylation-dependent manner. Furthermore, the Fluoppi assay showed that pathogenic mutations in these domains blocked interactions with phosphomimetic ubiquitin in mammalian cells.Thus molecular modeling based on the site-specific photo-crosslinking interaction map combined with mass spectrometry reveals that a novel binding mechanism between Parkin and ubiquitin.
All 2015 2014 Other
All Journal Article (6 results) (of which Peer Reviewed: 4 results, Acknowledgement Compliant: 1 results, Open Access: 2 results) Presentation (3 results) (of which Int'l Joint Research: 1 results, Invited: 3 results) Remarks (1 results)
The Journal of Biological Chemistry
Volume: 290 Issue: 42 Pages: 25199-25211
10.1074/jbc.m115.671446
The Journal of Cell Biology
Volume: 209 Issue: 1 Pages: 111-128
10.1083/jcb.201410050
J Cell Sci
Volume: 128 Pages: 964-978
10.1242/jcs.161000
Nature
Volume: 510(in press) Issue: 7503 Pages: 162-166
10.1038/nature13392
実験医学
Volume: 32 Pages: 2616-2620
細胞工学
Volume: 33 Pages: 974-976
http://www.igakuken.or.jp/project/detail/ubiquitin.html
