1WLC

Congerin II Y16S/T88I double mutant

1WLC の概要

• エントリーDOI: 10.2210/pdb1wlc/pdb
• 関連するPDBエントリー: 1IS3 1WLD
• 分子名称: Congerin II, 2-(N-MORPHOLINO)-ETHANESULFONIC ACID (3 entities in total)
• 機能のキーワード: galectin, beta-sandwich, thermostability, double-mutant, sugar binding protein
• 由来する生物種: Conger myriaster (whitespotted conger)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 15485.31

構造登録者

Shionyu-Mitsuyama, C.,Ito, Y.,Konno, A.,Miwa, Y.,Ogawa, T.,Muramoto, K.,Shirai, T. (登録日: 2004-06-22, 公開日: 2005-06-07, 最終更新日: 2024-05-29)

主引用文献

Shionyu-Mitsuyama, C.,Ito, Y.,Konno, A.,Miwa, Y.,Ogawa, T.,Muramoto, K.,Shirai, T.
In vitro evolutionary thermostabilization of congerin II: a limited reproduction of natural protein evolution by artificial selection pressure
J.Mol.Biol., 347:385-397, 2005

PubMed Abstract: The thermostability of the conger eel galectin, congerin II, was improved by in vitro evolutionary protein engineering. Two rounds of random PCR mutagenesis and selection experiments increased the congerin II thermostability to a level comparative to its naturally thermostable isoform, congerin I. The crystal structures of the most thermostable double mutant, Y16S/T88I, and the related single mutants, Y16S and T88I, were determined at 2.0 angstroms, 1.8 angstroms, and 1.6 angstroms resolution, respectively. The exclusion of two interior water molecules by the Thr88Ile mutation, and the relief of adjacent conformational stress by the Tyr16Ser mutation were the major contributions to the thermostability. These features in the congerin II mutants are similar to those observed in congerin I. The natural evolution of congerin genes, with the K(A)/K(S) ratio of 2.6, was accelerated under natural selection pressures. The thermostabilizing selection pressure artificially applied to congerin II mimicked the implied natural pressure on congerin I. The results showed that the artificial pressure made congerin II partially reproduce the natural evolution of congerin I.

PubMed: 15740748
DOI: 10.1016/j.jmb.2005.01.027

実験手法

X-RAY DIFFRACTION (2 Å)