Effect of an intersubunit disulfide bond on the stability of Streptomyces subtilisin inhibitor.


Abstract

The effect of an engineered disulfide bond between two identical subunits of a dimeric protein, Streptomyces subtilisin inhibitor, on the stability of the protein was studied by differential scanning calorimetry. The introduction of the linkage caused a large stabilization without changing the cooperativity of unfolding, with the denaturation temperature of a 2 mg/mL solution being increased by 14.3 degrees C to 95.0 degrees C at pH 9.5 and by 16.4 degrees C to 63.0 degrees C at pH 3.0. The stabilization was caused by a loss of denaturational entropy, i.e., -40 and -98 cal K-1 mol-1 at pH 3.0 and 9.5, respectively, which more than compensated for the loss in the denaturational enthalpy. Study holds ProTherm entries: 4041, 4042, 4043, 4044, 4045, 4046, 4047, 4048, 4049, 4050, 4051, 4052, 4053, 4054, 4055, 4056, 4057, 4058, 4059, 4060, 4061, 4062, 4063, 4064, 4065, 4066, 4067, 4068, 4069, 4070, 4071, 4072, 4073, 4074, 4075, 4076, 4077, 4078, 4079, 4080, 4081, 4082, 4083, 4084, 4085, 4086, 4087, 4088, 4089, 4090, 4091, 4092 Extra Details: engineered disulfide bond; dimeric protein;,cooperativity of unfolding; denaturational entropy; enthalpy

Submission Details

ID: Y8i4DBSU4

Submitter: Connie Wang

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

Version: 1

Publication Details
Tamura A;Kojima S;Miura K;Sturtevant JM,Biochemistry (1994) Effect of an intersubunit disulfide bond on the stability of Streptomyces subtilisin inhibitor. PMID:7981212
Additional Information

Study Summary

Number of data points 74
Proteins Subtilisin inhibitor ; STREPTOMYCES SUBTILISIN INHIBITOR
Unique complexes 3
Assays/Quantities/Protocols Experimental Assay: Tm pH:3.0 ; Experimental Assay: Tm pH:2.89 ; Experimental Assay: Tm pH:2.88 ; Experimental Assay: Tm pH:2.8 ; Experimental Assay: Tm pH:2.79 ; Experimental Assay: Tm pH:2.68 ; Experimental Assay: Tm pH:2.59 ; Experimental Assay: Tm pH:2.56 ; Experimental Assay: Tm pH:2.5 ; Experimental Assay: Tm pH:2.4 ; Experimental Assay: Tm pH:2.37 ; Experimental Assay: Tm pH:2.3 ; Experimental Assay: Tm pH:9.75 ; Experimental Assay: Tm pH:9.5 ; Experimental Assay: Tm pH:9.2 ; Experimental Assay: Tm buffers:phosphate: 25 mM, pH:7.0 ; Experimental Assay: Tm pH:3.25 ; Experimental Assay: Tm pH:3.24 ; Experimental Assay: Tm pH:3.21 ; Experimental Assay: Tm pH:3.2 ; Experimental Assay: Tm pH:3.19 ; Experimental Assay: Tm pH:3.16 ; Experimental Assay: Tm pH:3.15 ; Experimental Assay: Tm pH:3.12 ; Experimental Assay: Tm pH:3.07 ; Experimental Assay: Tm pH:3.04 ; Experimental Assay: Tm pH:3.03 ; Experimental Assay: Tm pH:2.98 ; Experimental Assay: Tm pH:2.94 ; Experimental Assay: Tm pH:2.9 ; Derived Quantity: dTm pH:3.21 ; Derived Quantity: dTm pH:3.2 ; Derived Quantity: dTm pH:3.19 ; Derived Quantity: dTm pH:2.98 ; Derived Quantity: dTm pH:2.9 ; Derived Quantity: dTm pH:9.75 ; Derived Quantity: dTm pH:9.5 ; Derived Quantity: dTm buffers:phosphate: 25 mM, pH:7.0 ; Derived Quantity: dTm pH:3.25 ; Derived Quantity: dTm pH:3.16 ; Derived Quantity: dTm pH:3.15 ; Derived Quantity: dTm pH:3.07 ; Derived Quantity: dTm pH:3.04 ; Derived Quantity: dTm pH:2.94
Libraries Mutations for sequence DAPSALYAPSALVLTVGKGVSATTAAPERAVTLTCAPGPSGTHPAAGSACADLAAVGGDLNALTRGEDVMCPMVYDPVLLTVDGVWQGKRVSYERVFSNECEMNAHGSSVFAF

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
3SSI 1996-03-01T00:00:00+0000 2.3 PROTEINASE INHIBITOR SSI (STREPTOMYCES SUBTILISIN, INHIBITOR) FROM STREPTOMYCES ALBOGRISEOLUS
3SIC 1991-08-30T00:00:00+0000 1.8 MOLECULAR RECOGNITION AT THE ACTIVE SITE OF SUBTILISIN BPN': CRYSTALLOGRAPHIC STUDIES USING GENETICALLY ENGINEERED PROTEINACEOUS INHIBITOR SSI (STREPTOMYCES SUBTILISIN INHIBITOR)
5SIC 1991-11-18T00:00:00+0000 2.2 MOLECULAR RECOGNITION AT THE ACTIVE SITE OF SUBTILISIN BPN': CRYSTALLOGRAPHIC STUDIES USING GENETICALLY ENGINEERED PROTEINACEOUS INHIBITOR SSI (STREPTOMYCES SUBTILISIN INHIBITOR)
2TLD 1991-09-16T00:00:00+0000 2.6 CRYSTAL STRUCTURE OF AN ENGINEERED SUBTILISIN INHIBITOR COMPLEXED WITH BOVINE TRYPSIN
2SIC 1991-04-01T00:00:00+0000 1.8 REFINED CRYSTAL STRUCTURE OF THE COMPLEX OF SUBTILISIN BPN' AND STREPTOMYCES SUBTILISIN INHIBITOR AT 1.8 ANGSTROMS RESOLUTION

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
90.7 Subtilisin inhibitor P28592 SSI_STRGI
100.0 Subtilisin inhibitor P01006 SSI_STRAO