6VJF

The P-Loop K to A mutation of C. therm Vps1 GTPase-BSE

6VJF の概要

• エントリーDOI: 10.2210/pdb6vjf/pdb
• 関連するPDBエントリー: 6DEF
• 分子名称: Putative sorting protein Vps1, PHOSPHOMETHYLPHOSPHONIC ACID GUANYLATE ESTER, MAGNESIUM ION, ... (4 entities in total)
• 機能のキーワード: gtpase, dynamin, dynamin superfamily, bse, ged, gcp, p-loop, hydrolase
• 由来する生物種: Chaetomium thermophilum (strain DSM 1495 / CBS 144.50 / IMI 039719); 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 173957.94

構造登録者

Tornabene, B.A.,Varlakhanova, N.V.,Chappie, J.S.,Ford, M.G.J. (登録日: 2020-01-15, 公開日: 2020-02-19, 最終更新日: 2023-10-11)

主引用文献

Tornabene, B.A.,Varlakhanova, N.V.,Hosford, C.J.,Chappie, J.S.,Ford, M.G.J.
Structural and functional characterization of the dominant negative P-loop lysine mutation in the dynamin superfamily protein Vps1.
Protein Sci., 29:1416-1428, 2020

PubMed Abstract: Dynamin-superfamily proteins (DSPs) are large self-assembling mechanochemical GTPases that harness GTP hydrolysis to drive membrane remodeling events needed for many cellular processes. Mutation to alanine of a fully conserved lysine within the P-loop of the DSP GTPase domain results in abrogation of GTPase activity. This mutant has been widely used in the context of several DSPs as a dominant-negative to impair DSP-dependent processes. However, the precise deficit of the P-loop K to A mutation remains an open question. Here, we use biophysical, biochemical and structural approaches to characterize this mutant in the context of the endosomal DSP Vps1. We show that the Vps1 P-loop K to A mutant binds nucleotide with an affinity similar to wild type but exhibits defects in the organization of the GTPase active site that explain the lack of hydrolysis. In cells, Vps1 and Dnm1 bearing the P-loop K to A mutation are defective in disassembly. These mutants become trapped in assemblies at the typical site of action of the DSP. This work provides mechanistic insight into the widely-used DSP P-loop K to A mutation and the basis of its dominant-negative effects in the cell.

PubMed: 31981262
DOI: 10.1002/pro.3830

実験手法

X-RAY DIFFRACTION (2.472 Å)