2VEL

Structure-based enzyme engineering efforts with an inactive monomeric TIM variant: the importance of a single point mutation for generating an active site with suitable binding properties.

2VEL の概要

• エントリーDOI: 10.2210/pdb2vel/pdb
• 関連するPDBエントリー: 1AG1 1DKW 1IIG 1IIH 1KV5 1ML1 1MSS 1MTM 1TPD 1TPE 1TPF 1TRD 1TRI 1TSI 1TTI 1TTJ 2J24 2J27 2V0T 2V2C 2V2D 2V2H 2V5L 2VEI 2VEK 3TIM 4TIM 5TIM 6TIM
• 分子名称: GLYCOSOMAL TRIOSEPHOSPHATE ISOMERASE, CHLORIDE ION, 2-PHOSPHOGLYCOLIC ACID, ... (4 entities in total)
• 機能のキーワード: isomerase, triosephosphate isomerase, tim barrel, glycolysis, engineering, pentose shunt, binding pocket, gluconeogenesis, lipid synthesis, substrate specificity, fatty acid biosynthesis, tim, enzyme, monomeric, glycosome
• 由来する生物種: TRYPANOSOMA BRUCEI BRUCEI
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 51701.55

構造登録者

Alahuhta, M.,Salin, M.,Casteleijn, M.G.,Kemmer, C.,El-Sayed, I.,Augustyns, K.,Neubauer, P.,Wierenga, R.K. (登録日: 2007-10-24, 公開日: 2008-02-19, 最終更新日: 2024-05-01)

主引用文献

Alahuhta, M.,Salin, M.,Casteleijn, M.G.,Kemmer, C.,El-Sayed, I.,Augustyns, K.,Neubauer, P.,Wierenga, R.K.
Structure-Based Protein Engineering Efforts with a Monomeric Tim Variant: The Importance of a Single Point Mutation for Generating an Active Site with Suitable Binding Properties.
Protein Eng.Des.Sel., 21:257-, 2008

PubMed Abstract: A monomeric variant of triosephosphate isomerase (TIM) with a new engineered binding groove has been characterized further. In this variant (ml8bTIM), the phosphate binding loop had been shortened, causing the binding site to be much more extended. Here, it is reported that in the V233A variant of ml8bTIM (A-TIM), three important properties of the wild-type TIM active site have been restored: (i) the structural properties of loop-7, (ii) the binding site of a conserved water molecule between loop-7 and loop-8 and (iii) the binding site of the phosphate moiety. It is shown that the active site of A-TIM can bind TIM transition state analogs and suicide inhibitors competently. It is found that the active site geometry of the A-TIM complexes is less compact and more solvent exposed, as in wild-type TIM. This correlates with the observation that the catalytic efficiency of A-TIM for interconverting the TIM substrates is too low to be detected. It is also shown that the A-TIM active site can bind compounds which do not bind to wild-type TIM and which are completely different from the normal TIM substrate, like a citrate molecule. The binding of this citrate molecule is stabilized by hydrogen bonding interactions with the new binding groove.

PubMed: 18239072
DOI: 10.1093/PROTEIN/GZN002

実験手法

X-RAY DIFFRACTION (2.3 Å)