Folding defects caused by single amino acid substitutions in a subunit are not alleviated by assembly.


Abstract

Significant stabilization of a protein often occurs when it is assembled into an oligomer. Bacteriophage P22 contains 420 monomers of coat protein that are stabilized by the assembly and maturation processes. The effects of eight single amino acid substitutions in coat protein that each cause a temperature-sensitive-folding defect were investigated to determine if the conformational differences previously observed in the monomers could be alleviated by assembly or maturation. Several techniques including differential scanning calorimetry, heat-induced expansion, urea denaturation, and sensitivity to protease digestion were used to explore the effects of the amino acid substitutions on the conformation of coat protein, once assembled. Each of the amino acid substitutions caused a change in the conformation as compared to wild-type coat protein, observed by at least one of the probes used. Thus, neither assembly nor expansion entirely corrected the conformational defects in the monomeric subunits of the folding mutants. Study holds ProTherm entries: 7995, 7996, 7997, 7998, 7999, 8000, 8001, 8002, 8003 Extra Details: additive : EDTA(2 mM), coat protein; temperature-sensitive-folding defect;,conformational differences

Submission Details

ID: B7e9pTtD4

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:35 p.m.

Version: 1

Publication Details
Capen CM;Teschke CM,Biochemistry (2000) Folding defects caused by single amino acid substitutions in a subunit are not alleviated by assembly. PMID:10653661
Additional Information

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
2M5S 2014-03-05 High-resolution NMR structure and cryo-EM imaging support multiple functional roles for the accessory I-domain of phage P22 coat protein
5UU5 2017-03-15 3.3 Bacteriophage P22 mature virion capsid protein
2XYY 2011-02-02 3.8 De Novo model of Bacteriophage P22 procapsid coat protein
2XYZ 2011-02-02 4.0 De Novo model of Bacteriophage P22 virion coat protein
3IYH 2010-03-31 8.2 P22 procapsid coat protein structures reveal a novel mechanism for capsid maturation: Stability without auxiliary proteins or chemical cross-links
3IYI 2010-03-31 9.1 P22 expanded head coat protein structures reveal a novel mechanism for capsid maturation: Stability without auxiliary proteins or chemical cross-links

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Major capsid protein P26747 CAPSD_BPP22