Stability enhancement of cytochrome c through heme deprotonation and mutations.


Abstract

The chemical denaturation of Pseudomonas aeruginosa cytochrome c(551) variants was examined at pH 5.0 and 3.6. All variants were stabilized at both pHs compared with the wild-type. Remarkably, the variants carrying the F34Y and/or E43Y mutations were more stabilized than those having the F7A/V13M or V78I ones at pH 5.0 compared with at pH 3.6 by ~3.0-4.6 kJ/mol. Structural analyses predicted that the side chains of introduced Tyr-34 and Tyr-43 become hydrogen donors for the hydrogen bond formation with heme 17-propionate at pH 5.0, but less efficiently at pH 3.6, because the propionate is deprotonated at the higher pH. Our results provide an insight into a stabilization strategy for heme proteins involving variation of the heme electronic state and introduction of appropriate mutations. Study holds ProTherm entries: 24920, 24921, 24922, 24923, 24924, 24925, 24926, 24927, 24928, 24929, 24930, 24931, 24932, 24933, 24934, 24935, 24936, 24937, 24938, 24939 Extra Details: Circular dichroism spectroscopy; Cytochrome c; Hydrogen bond; Protein stability; Heme propionate

Submission Details

ID: FB65dEC54

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:55 p.m.

Version: 1

Publication Details
Sonoyama T;Hasegawa J;Uchiyama S;Nakamura S;Kobayashi Y;Sambongi Y,Biophys. Chem. (2009) Stability enhancement of cytochrome c through heme deprotonation and mutations. PMID:18957274
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
2PAC 1993-10-31 SOLUTION STRUCTURE OF FE(II) CYTOCHROME C551 FROM PSEUDOMONAS AERUGINOSA AS DETERMINED BY TWO-DIMENSIONAL 1H NMR
5XEC 2017-08-09 1.1 Heterodimer constructed from PA cyt c551-HT cyt c552 and HT cyt c552-PA cyt c551 chimeric proteins
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
2EXV 2006-02-07 1.86 Crystal structure of the F7A mutant of the cytochrome c551 from Pseudomonas aeruginosa

Relevant UniProtKB Entries

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