Thermodynamic characterization of variants of mesophilic cytochrome c and its thermophilic counterpart.


Abstract

Thermal stability was measured for variants of cytochrome c-551 (PA c-551) from a mesophile, Pseudomonas aeruginosa, and a thermophilic counterpart, Hydrogenobacter thermophilus cytochrome c-552 (HT c-552), by differential scanning calorimetry (DSC) at pH 3.6. The mutated residues in PA c-551, selected with reference to the corresponding residues in HT c-552, were located in three spatially separated regions: region I, Phe7 to Ala/Val13 to Met; region II, Glu34 to Tyr/Phe43 to Tyr; and region III, Val78 to Ile. The thermodynamic parameters determined indicated that the mutations in regions I and III caused enhanced stability through not only enthalpic but also entropic contributions, which reflected improved packing of the side chains. Meanwhile, the mutated region II made enthalpic contributions to the stability through electrostatic interactions. The obtained differences in the Gibbs free energy changes of unfolding [Delta(DeltaG)] showed that the three regions contributed to the overall stability in an additive manner. HT c-552 had the smallest heat capacity change (DeltaC(P)), resulting in higher DeltaG values over a wide temperature range (0-100 degrees C), compared to the PA c-551 variants; this contributed to the highest stability of HT c-552. Our DSC measurement results, in conjunction with mutagenesis and structural studies on the homologous mesophilic and thermophilic cytochromes c, provided an extended thermodynamic view of protein stabilization. Study holds ProTherm entries: 15177, 15178, 15179, 15180, 15181, 15182, 15183, 15184, 15185, 15186, 15187, 15188, 15189, 15190, 15191, 15192, 15193, 15194 Extra Details: combined mutant; cytochrome c; differential scanning calorimetry (DSC); protein stability

Submission Details

ID: t3kUTbrn3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:45 p.m.

Version: 1

Publication Details
Uchiyama S;Hasegawa J;Tanimoto Y;Moriguchi H;Mizutani M;Igarashi Y;Sambongi Y;Kobayashi Y,Protein Eng. (2002) Thermodynamic characterization of variants of mesophilic cytochrome c and its thermophilic counterpart. PMID:12082163
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
2PAC 1993-10-31 SOLUTION STRUCTURE OF FE(II) CYTOCHROME C551 FROM PSEUDOMONAS AERUGINOSA AS DETERMINED BY TWO-DIMENSIONAL 1H NMR
1DVV 2000-11-29 SOLUTION STRUCTURE OF THE QUINTUPLE MUTANT OF CYTOCHROME C-551 FROM PSEUDOMONAS AERUGINOSA
2AI5 2006-05-23 Solution Structure of Cytochrome C552, determined by Distributed Computing Implementation for NMR data
1AYG 1998-11-25 SOLUTION STRUCTURE OF CYTOCHROME C-552, NMR, 20 STRUCTURES
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