The role of aromatic residues in the hydrophobic core of the villin headpiece subdomain.


Abstract

Small autonomously folding proteins are of interest as model systems to study protein folding, as the same molecule can be used for both experimental and computational approaches. The question remains as to how well these minimized peptide model systems represent larger native proteins. For example, is the core of a minimized protein tolerant to mutation like larger proteins are? Also, do minimized proteins use special strategies for specifying and stabilizing their folded structure? Here we examine these questions in the 35-residue autonomously folding villin headpiece subdomain (VHP subdomain). Specifically, we focus on a cluster of three conserved phenylalanine (F) residues F47, F51, and F58, that form most of the hydrophobic core. These three residues are oriented such that they may provide stabilizing aromatic-aromatic interactions that could be critical for specifying the fold. Circular dichroism and 1D-NMR spectroscopy show that point mutations that individually replace any of these three residues with leucine were destabilized, but retained the native VHP subdomain fold. In pair-wise replacements, the double mutant that retains F58 can adopt the native fold, while the two double mutants that lack F58 cannot. The folding of the double mutant that retains F58 demonstrates that aromatic-aromatic interactions within the aromatic cluster are not essential for specifying the VHP subdomain fold. The ability of the VHP subdomain to tolerate mutations within its hydrophobic core indicates that the information specifying the three dimensional structure is distributed throughout the sequence, as observed in larger proteins. Thus, the VHP subdomain is a legitimate model for larger, native proteins. Study holds ProTherm entries: 12998, 12999, 13000, 13001, 13002, 13003, 13004, 13005 Extra Details: villin headpiece subdomain; protein folding; aromatic-aromatic,interactions; hydrophobic core mutation

Submission Details

ID: Rn3vhB6T3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:44 p.m.

Version: 1

Publication Details
Frank BS;Vardar D;Buckley DA;McKnight CJ,Protein Sci. (2002) The role of aromatic residues in the hydrophobic core of the villin headpiece subdomain. PMID:11847290
Additional Information

Study Summary

Number of data points 8
Proteins Villin-1 ; Villin-1
Unique complexes 8
Assays/Quantities/Protocols Experimental Assay: Tm
Libraries Mutations for sequence PTKLETFPLDVLVNTAAEDLPRGVDPSRKENHLSDEDFKAVFGMTRSAFANLPLWKQQNLKKEKGLF

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
2JM0 2006-10-17 Solution structure of chicken villin headpiece subdomain containing a fluorinated side chain in the core
2VIL 1997-04-01 REFINED STRUCTURE OF THE ACTIN-SEVERING DOMAIN VILLIN 14T, DETERMINED BY SOLUTION NMR, 11 STRUCTURES
1VII 1997-08-12 THERMOSTABLE SUBDOMAIN FROM CHICKEN VILLIN HEADPIECE, NMR, MINIMIZED AVERAGE STRUCTURE
2LLF 2012-11-07 Sixth Gelsolin-like domain of villin in 5 mM CaCl2
4CZ4 2015-02-18 HP24stab derived from the villin headpiece subdomain
4CZ3 2015-02-18 HP24wt derived from the villin headpiece subdomain
1QQV 1999-12-29 SOLUTION STRUCTURE OF THE HEADPIECE DOMAIN OF CHICKEN VILLIN
2VIK 1997-04-01 REFINED STRUCTURE OF THE ACTIN-SEVERING DOMAIN VILLIN 14T, DETERMINED BY SOLUTION NMR, MINIMIZED AVERAGE STRUCTURE
2PPZ 2007-05-29 NMR solution Structure of the Villin Headpiece Mutant G34L
1WY3 2005-05-03 0.95 Chicken villin subdomain HP-35, K65(NLE), N68H, pH7.0
3TRV 2012-01-25 1.0 Crystal structure of quasiracemic villin headpiece subdomain containing (F5Phe17) substitution
1YRI 2005-05-03 1.0 Chicken villin subdomain HP-35, N68H, pH6.4
2F4K 2006-04-11 1.05 Chicken villin subdomain HP-35, K65(NLE), N68H, K70(NLE), PH9
1YRF 2005-05-03 1.07 Chicken villin subdomain HP-35, N68H, pH6.7
5I1S 2016-05-25 1.12 Villin headpiece subdomain with a Lys30 to APC substitution
5I1N 2016-05-25 1.3 Villin headpiece subdomain with a Gln26 to beta-3-homoglutamine substitution
5I1O 2016-05-25 1.35 Villin headpiece subdomain with a Gln26 to ACPC substitution
2RJY 2008-09-23 1.4 Crystal structure of villin headpiece, P21 21 21 space group
1YU5 2005-09-06 1.4 Crystal Structure of the Headpiece Domain of Chicken Villin
5I1P 2016-05-25 1.4 Villin headpiece subdomain with a Lys30 to beta-3-homolysine substitution
2RJV 2008-09-23 1.45 Crystal structure of the H41Y mutant of villin headpiece, P 21 21 21 space group
1YU8 2005-09-06 1.45 Crystal Structure of the R37A Mutant of Villin Headpiece
3TJW 2012-01-25 1.46 Crystal Structure of Quasiracemic Villin Headpiece Subdomain Containing (F5Phe10) Substitution
1YU7 2005-09-06 1.5 Crystal Structure of the W64Y mutant of Villin Headpiece
2RJW 2009-02-17 1.55 The crystal structure of the H41Y mutant of villin headpiece, P61 SPACE GROUP.
1WY4 2005-05-03 1.55 Chicken villin subdomain HP-35, K65(NLE), N68H, pH5.1
3NKJ 2011-06-01 1.6 Crystal Structure of HP67 L61G
2RJX 2009-02-17 1.7 Crystal structure of the headpiece domain of chicken villin, P61 space group
3MYC 2011-11-02 1.7 Crystal Structure of HP67 H41F - P212121
3MYE 2011-05-25 1.8 Crystal Structure of HP67 L61GL
3TRW 2012-01-25 2.1 Crystal structure of racemic villin headpiece subdomain crystallized in space group P-1
3TRY 2012-01-25 2.3 Crystal structure of racemic villin headpiece subdomain in space group I-4c2
3MYA 2011-11-02 2.5 Crystal Structure of HP67 H41F - P61

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Villin-1 P02640 VILI_CHICK