Selected mutations in a mesophilic cytochrome c confer the stability of a thermophilic counterpart.


Abstract

Mesophilic cytochrome c(551) of Pseudomonas aeruginosa (PA c(551)) became as stable as its thermophilic counterpart, Hydrogenobacter thermophilus cytochrome c(552) (HT c(552)), through only five amino acid substitutions. The five residues, distributed in three spatially separated regions, were selected and mutated with reference to the corresponding residues in HT c(552) through careful structure comparison. Thermodynamic analysis indicated that the stability of the quintuple mutant of PA c(551) could be partly attained through an enthalpic factor. The solution structure of the mutant showed that, as in HT c(552), there were tighter side chain packings in the mutated regions. Furthermore, the mutant had an increased total accessible surface area, resulting in great negative hydration free energy. Our results provide a novel example of protein stabilization in that limited amino acid substitutions can confer the overall stability of a natural highly thermophilic protein upon a mesophilic molecule. Study holds ProTherm entries: 9769, 9770, 9771, 9772, 9773, 9774, 14512, 14513 Extra Details: mesophilic; thermophilic; structure comparison; quintuple mutant;,side chain packings; hydration free energy

Submission Details

ID: QqeaeiF7

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:38 p.m.

Version: 1

Publication Details
Hasegawa J;Uchiyama S;Tanimoto Y;Mizutani M;Kobayashi Y;Sambongi Y;Igarashi Y,J. Biol. Chem. (2000) Selected mutations in a mesophilic cytochrome c confer the stability of a thermophilic counterpart. PMID:10918067
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
1DVV 2000-11-29 SOLUTION STRUCTURE OF THE QUINTUPLE MUTANT OF CYTOCHROME C-551 FROM PSEUDOMONAS AERUGINOSA
1AYG 1998-11-25 SOLUTION STRUCTURE OF CYTOCHROME C-552, NMR, 20 STRUCTURES
2PAC 1993-10-31 SOLUTION STRUCTURE OF FE(II) CYTOCHROME C551 FROM PSEUDOMONAS AERUGINOSA AS DETERMINED BY TWO-DIMENSIONAL 1H NMR
2AI5 2006-05-23 Solution Structure of Cytochrome C552, determined by Distributed Computing Implementation for NMR data
5XEC 2017-08-09 1.1 Heterodimer constructed from PA cyt c551-HT cyt c552 and HT cyt c552-PA cyt c551 chimeric proteins
5AUR 2015-10-21 1.26 Hydrogenobacter thermophilus cytochrome c552 dimer formed by domain swapping at N-terminal region
5AUS 2015-10-21 1.3 Hydrogenobacter thermophilus cytochrome c552 dimer formed by domain swapping at C-terminal region
3X39 2015-04-22 1.5 Domain-swapped dimer of Pseudomonas aeruginosa cytochrome c551
5XED 2017-08-09 1.55 Heterodimer constructed from M61A PA cyt c551-HT cyt c552 and HT cyt c552-PA cyt c551 chimeric proteins
451C 1981-10-02 1.6 STRUCTURE OF CYTOCHROME C551 FROM P. AERUGINOSA REFINED AT 1.6 ANGSTROMS RESOLUTION AND COMPARISON OF THE TWO REDOX FORMS
351C 1981-10-02 1.6 STRUCTURE OF CYTOCHROME C551 FROM P. AERUGINOSA REFINED AT 1.6 ANGSTROMS RESOLUTION AND COMPARISON OF THE TWO REDOX FORMS
4ZID 2016-02-10 1.8 Dimeric Hydrogenobacter thermophilus cytochrome c552 obtained from Escherichia coli
2EXV 2006-02-07 1.86 Crystal structure of the F7A mutant of the cytochrome c551 from Pseudomonas aeruginosa
3VYM 2012-11-07 2.0 Dimeric Hydrogenobacter thermophilus cytochrome c552
1YNR 2005-05-17 2.0 Crystal structure of the cytochrome c-552 from Hydrogenobacter thermophilus at 2.0 resolution

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Cytochrome c-552 P15452 CY552_HYDTT
100.0 Cytochrome c-551 P00099 CY551_PSEAE