9B6C

Human asparagine synthetase Arg-142 to Ile-142 (R142I) variant

9B6C の概要

• エントリーDOI: 10.2210/pdb9b6c/pdb
• EMDBエントリー: 40764 44253
• 分子名称: Asparagine synthetase [glutamine-hydrolyzing] (1 entity in total)
• 機能のキーワード: asparagine, aspartic acid, glutamine, ammonia, biosynthetic protein
• 由来する生物種: Homo sapiens (human)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 133344.23

構造登録者

Coricello, A.,Nardone, A.,Lupia, A.,Gratteri, C.,Vos, M.,Chaptal, V.,Alcaro, S.,Zhu, W.,Takagi, Y.,Richards, N. (登録日: 2024-03-25, 公開日: 2025-01-08, 最終更新日: 2025-02-05)

主引用文献

Coricello, A.,Nardone, A.J.,Lupia, A.,Gratteri, C.,Vos, M.,Chaptal, V.,Alcaro, S.,Zhu, W.,Takagi, Y.,Richards, N.G.J.
3D variability analysis reveals a hidden conformational change controlling ammonia transport in human asparagine synthetase.
Nat Commun, 15:10538-10538, 2024

PubMed Abstract: Advances in X-ray crystallography and cryogenic electron microscopy (cryo-EM) offer the promise of elucidating functionally relevant conformational changes that are not easily studied by other biophysical methods. Here we show that 3D variability analysis (3DVA) of the cryo-EM map for wild-type (WT) human asparagine synthetase (ASNS) identifies a functional role for the Arg-142 side chain and test this hypothesis experimentally by characterizing the R142I variant in which Arg-142 is replaced by isoleucine. Support for Arg-142 playing a role in the intramolecular translocation of ammonia between the active site of the enzyme is provided by the glutamine-dependent synthetase activity of the R142 variant relative to WT ASNS, and MD simulations provide a possible molecular mechanism for these findings. Combining 3DVA with MD simulations is a generally applicable approach to generate testable hypotheses of how conformational changes in buried side chains might regulate function in enzymes.

PubMed: 39627226
DOI: 10.1038/s41467-024-54912-9

実験手法

ELECTRON MICROSCOPY (3.35 Å)