2K6N

Solution Structure of Human Supervillin Headpiece, Minimized Average

Summary for 2K6N

• Entry DOI: 10.2210/pdb2k6n/pdb
• Related: 1QQV 1QZP 1UJS 1UNC 1UND 1VII 1WY3 1WY4 1YRF 1YRI 1YU5 1YU7 1YU8 1ZV6 2JM0 2K6M 2PPZ
• NMR Information: BMRB: 15874
• Descriptor: Supervillin (1 entity in total)
• Functional Keywords: supervillin, svhp, hp, headpiece, villin, archvillin, actin capping, actin-binding, alternative splicing, calcium, cytoplasm, cytoskeleton, membrane, phosphoprotein, polymorphism, structural protein
• Biological source: Homo sapiens (human)
• Cellular location: Cell membrane; Peripheral membrane protein; Cytoplasmic side: O95425
• Total number of polymer chains: 1
• Total formula weight: 7708.04

Authors

Brown, J.W.,Didem, V.,McKnight, C. (deposition date: 2008-07-11, release date: 2009-07-14, Last modification date: 2024-05-29)

Primary citation

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

Experimental method

SOLUTION NMR