1M9I

Crystal Structure Of Phosphorylation-Mimicking Mutant T356D Of Annexin VI

1M9I の概要

• エントリーDOI: 10.2210/pdb1m9i/pdb
• 関連するPDBエントリー: 1AVC
• 分子名称: Annexin VI, CALCIUM ION (3 entities in total)
• 機能のキーワード: annexin, calcium-binding, membrane-binding, phosphorylation, mutant t356d, lipid binding protein
• 由来する生物種: Homo sapiens (human)
• 細胞内の位置: Cytoplasm (By similarity): P08133
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 76049.44

構造登録者

Freye-Minks, C.,Kretsinger, R.H.,Creutz, C.E. (登録日: 2002-07-29, 公開日: 2002-08-07, 最終更新日: 2024-04-03)

主引用文献

Freye-Minks, C.,Kretsinger, R.H.,Creutz, C.E.
Structural and Dynamic Changes in Human Annexin VI Induced by a Phosphorylation-Mimicking Mutation, T356D
Biochemistry, 42:620-630, 2003

PubMed Abstract: Phosphorylation of some members of the annexin family of proteins may play a significant role in controlling their calcium-dependent interactions with membranes. Recent electron microscopic studies of annexin VI revealed that the protein's two core domains exhibit a great degree of flexibility and are able to undergo a relative conformational change that could potentially initiate contacts between membranes [Avila-Sakar, A. J., et al. (2000) J. Struct. Biol. 130, 54-62]. To assess the possibility of a regulatory role of phosphorylation in this behavior, the crystal structure of a phosphorylation-mimicking mutant (T356D in the flexible connector region of human annexin VI) was determined to 2.65 A resolution. When the mutant is compared to the wild-type annexin VI, subtle differences are seen at the site of the mutation, while larger changes are evident in one of the calcium-binding loops and in the presence of five calcium ions. Furthermore, biochemical studies provide evidence for additional conformational differences between the T356D and wild-type solution structures. Fluorescence emission and acrylamide quenching suggest a higher level of solvent exposure of Trp-343 in the connector region of T356D in the presence of calcium. Comparisons of retardation coefficients in native gel electrophoresis reveal that T356D has a more extended shape, while proteolytic studies show a greater accessibility of a trypsin cleavage site inside the linker region, indicating a conformation more open than the wild-type form. These data provide insights into a possible regulatory mechanism leading to a higher degree of flexibility and possibly a higher calcium binding affinity of annexin VI upon phosphorylation.

PubMed: 12534274
DOI: 10.1021/bi026742h

実験手法

X-RAY DIFFRACTION (2.65 Å)