1BCZ

RECOMBINANT RAT ANNEXIN V, T72S MUTANT

1BCZ の概要

• エントリーDOI: 10.2210/pdb1bcz/pdb
• 分子名称: ANNEXIN V, CALCIUM ION, SULFATE ION, ... (4 entities in total)
• 機能のキーワード: calcium binding protein, phospholipid membrane binding protein, calcium-binding protein
• 由来する生物種: Rattus norvegicus (Norway rat)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 36070.89

構造登録者

Mo, Y.D.,Swairjo, M.A.,Li, C.W.,Head, J.F.,Seaton, B.A. (登録日: 1998-05-04, 公開日: 1998-11-25, 最終更新日: 2024-02-07)

主引用文献

Campos, B.,Mo, Y.D.,Mealy, T.R.,Li, C.W.,Swairjo, M.A.,Balch, C.,Head, J.F.,Retzinger, G.,Dedman, J.R.,Seaton, B.A.
Mutational and crystallographic analyses of interfacial residues in annexin V suggest direct interactions with phospholipid membrane components.
Biochemistry, 37:8004-8010, 1998

PubMed Abstract: Annexin V belongs to a family of eukaryotic calcium-dependent membrane-binding proteins. The calcium-binding sites at the annexin-membrane interface have been investigated in some detail; however, little is known about the functional roles of highly conserved interfacial residues that do not coordinate calcium themselves. In the present study, the importance of tryptophan 185, and threonine or serine at positions 72, 144, 228, and 303, in rat annexin V is investigated by site-directed mutagenesis, X-ray crystallography, and functional assays. The high-resolution crystal structures of the mutants show that the mutations do not cause structural perturbations of the annexin molecule itself or disappearance of bound calcium ions from calcium-binding sites. The assays indicate that relative to wild-type annexin V, loss of the methyl substituent at position 72 (Thr72-->Ser) has no effect while loss of the hydroxyl group (Thr72-->Ala or Thr72-->Lys) causes reduction of membrane binding. Multiple lysine substitutions (e.g., Thr72,Ser144,Ser228,Ser303-->Lys) have a greater adverse effect than the single lysine mutation, suggesting that in annexin V the introduction of potentially favorable electrostatic interactions between the lysine side chains and the net negatively charged membrane surface is not sufficient to overcome the loss of the hydroxyl side chains. Replacement of the unique tryptophan, Trp185, by alanine similarly decreases membrane binding affinity. Taken together, the data suggest that the side chains mutated in this study contribute to phospholipid binding and participate directly in intermolecular contacts with phospholipid membrane components.

PubMed: 9609693
DOI: 10.1021/bi973142n

実験手法

X-RAY DIFFRACTION (2.2 Å)