2K6M

Solution Structure of Human Supervillin Headpiece

2K6M の概要

• エントリーDOI: 10.2210/pdb2k6m/pdb
• 関連するPDBエントリー: 1QQV 1QZP 1UJS 1UNC 1UND 1VII 1WY3 1WY4 1YRF 1YRI 1YU5 1YU7 1YU8 1ZV6 2JM0 2K6N 2PPZ
• NMR情報: BMRB: 15873
• 分子名称: Supervillin (1 entity in total)
• 機能のキーワード: supervillin, svhp, hp, headpiece, villin, archvillin, actin capping, actin-binding, alternative splicing, calcium, cytoplasm, cytoskeleton, membrane, phosphoprotein, polymorphism, structural protein
• 由来する生物種: Homo sapiens (human)
• 細胞内の位置: Cell membrane; Peripheral membrane protein; Cytoplasmic side: O95425
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 7708.04

構造登録者

Brown, J.W.,Didem, V.,McKnight, C. (登録日: 2008-07-11, 公開日: 2009-07-14, 最終更新日: 2024-05-29)

主引用文献

Brown, J.W.,Vardar-Ulu, D.,McKnight, C.J.
How to arm a supervillin: designing F-actin binding activity into supervillin headpiece.
J.Mol.Biol., 393:608-618, 2009

PubMed Abstract: Villin-type headpiece domains are compact motifs that have been used extensively as model systems for protein folding. Although the majority of headpiece domains bind actin, there are some that lack this activity. Here, we present the first NMR solution structure and (15)N-relaxation analysis of a villin-type headpiece domain natively devoid of F-actin binding activity, that of supervillin headpiece (SVHP). The structure was found to be similar to that of other headpiece domains that bind F-actin. Our NMR analysis demonstrates that SVHP lacks a conformationally flexible region (V-loop) present in all other villin-type headpiece domains and which is essential to the phosphoryl regulation of dematin headpiece. In comparing the electrostatic surface potential map of SVHP to that of other villin-type headpiece domains with significant affinity for F-actin, we identified a positive surface potential conserved among headpiece domains that bind F-actin but absent from SVHP. A single point mutation (L38K) in SVHP, which creates a similar positive surface potential, endowed SVHP with specific affinity for F-actin that is within an order of magnitude of the tightest binding headpiece domains. We propose that this effect is likely conferred by a specific buried salt bridge between headpiece and actin. As no high-resolution structural information exists for the villin-type headpiece F-actin complex, our results demonstrate that through positive mutagenesis, it is possible to design binding activity into homologous proteins without structural information of the counterpart's binding surface.

PubMed: 19683541
DOI: 10.1016/j.jmb.2009.08.018

実験手法

SOLUTION NMR