Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 116, Issue 31, Pages 15475-15484Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1905488116
Keywords
ubiquitin; adenylation; thioester; X-ray; E1
Categories
Funding
- NIH National Institute of General Medical Sciences (NIGMS) [R01 GM065872, R35 GM118080, R01 GM100477]
- NIH National Cancer Institute-Cancer Center Support Grant [P30 CA008748]
- NSF Graduate Research Fellowship [2015190598]
- NIH Chemistry-Biology Interface Training Grant [T32 GM115327]
- NIH NIGMS Grant [P41 GM103403]
- NIH Office of Research Infrastructure Programs High-End Instrumentation Grant [S10 RR029205]
- DOE Office of Science [DE-AC02-06CH11357]
Ask authors/readers for more resources
The ubiquitin (Ub) and Ub-like (Ubl) protein-conjugation cascade is initiated by E1 enzymes that catalyze Ub/Ubl activation through C-terminal adenylation, thioester bond formation with an E1 catalytic cysteine, and thioester bond transfer to Ub/Ubl E2 conjugating enzymes. Each of these reactions is accompanied by conformational changes of the E1 domain that contains the catalytic cysteine (Cys domain). Open conformations of the Cys domain are associated with adenylation and thioester transfer to E2s, while a closed conformation is associated with pyrophosphate release and thioester bond formation. Several structures are available for Ub E1s, but none has been reported in the open state before pyrophosphate release or in the closed state. Here, we describe the structures of Schizosaccharomyces pombe Ub E1 in these two states, captured using semisynthetic Ub probes. In the first, with a Ub-adenylate mimetic (Ub-AMSN) bound, the E1 is in an open conformation before release of pyrophosphate. In the second, with a Ub-vinylsulfonamide (Ub-AVSN) bound covalently to the catalytic cysteine, the E1 is in a closed conformation required for thioester bond formation. These structures provide further insight into Ub E1 adenylation and thioester bond formation. Conformational changes that accompany Cys-domain rotation are conserved for SUMO and Ub E1s, but changes in Ub E1 involve additional surfaces as mutational and biochemical analysis of residues within these surfaces alter Ub E1 activities.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available