5SUL

Inhibited state structure of yGsy2p

5SUL の概要

• エントリーDOI: 10.2210/pdb5sul/pdb
• 分子名称: Glycogen [starch] synthase isoform 2, URIDINE-5'-DIPHOSPHATE, URIDINE-5'-MONOPHOSPHATE (3 entities in total)
• 機能のキーワード: glycogen synthase inhibited state, phosphorylation, transferase
• 由来する生物種: Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (Baker's yeast)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 165126.08

構造登録者

Mahalingan, K.K.,Hurley, T.D. (登録日: 2016-08-03, 公開日: 2017-06-14, 最終更新日: 2023-10-04)

主引用文献

Mahalingan, K.K.,Baskaran, S.,DePaoli-Roach, A.A.,Roach, P.J.,Hurley, T.D.
Redox Switch for the Inhibited State of Yeast Glycogen Synthase Mimics Regulation by Phosphorylation.
Biochemistry, 56:179-188, 2017

PubMed Abstract: Glycogen synthase (GS) is the rate limiting enzyme in the synthesis of glycogen. Eukaryotic GS is negatively regulated by covalent phosphorylation and allosterically activated by glucose-6-phosphate (G-6-P). To gain structural insights into the inhibited state of the enzyme, we solved the crystal structure of yGsy2-R589A/R592A to a resolution of 3.3 Å. The double mutant has an activity ratio similar to the phosphorylated enzyme and also retains the ability to be activated by G-6-P. When compared to the 2.88 Å structure of the wild-type G-6-P activated enzyme, the crystal structure of the low-activity mutant showed that the N-terminal domain of the inhibited state is tightly held against the dimer-related interface thereby hindering acceptor access to the catalytic cleft. On the basis of these two structural observations, we developed a reversible redox regulatory feature in yeast GS by substituting cysteine residues for two highly conserved arginine residues. When oxidized, the cysteine mutant enzyme exhibits activity levels similar to the phosphorylated enzyme but cannot be activated by G-6-P. Upon reduction, the cysteine mutant enzyme regains normal activity levels and regulatory response to G-6-P activation.

PubMed: 27935293
DOI: 10.1021/acs.biochem.6b00884

実験手法

X-RAY DIFFRACTION (3.3 Å)