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

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
Sequence Assay Result Units