Folding and stability of a fibronectin type III domain of human tenascin.


Abstract

The folding of an isolated fibronectin type III domain of human tenascin, a large extra-cellular matrix protein, has been characterised. The isolated module, which has no disulphide bonds, can be reversibly unfolded by chemical denaturant and temperature. Equilibrium unfolding, measured using a number of different probes, fits to a two-state transition, with consistent measures of DeltaGH2OD-N. Folding and refolding rate constants have been determined over a range of denaturant concentrations. The refolding kinetics are bi-phasic, and in the transition region the slow phase dominates refolding kinetics. Outside the transition region the folding of the fast-folding species fits to a two-state model. There is no evidence for significant accumulation of partially folded intermediates. Study holds ProTherm entries: 8887, 8888, 8889, 8890, 8891, 8892, 8893, 8894, 8895, 8896 Extra Details: Fluorescence measurements at 350nm tenascin; fibronectin type III; immunoglobulin; protein folding; stability

Submission Details

ID: xfkYQ7rA4

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:37 p.m.

Version: 1

Publication Details
Clarke J;Hamill SJ;Johnson CM,J. Mol. Biol. (1997) Folding and stability of a fibronectin type III domain of human tenascin. PMID:9245604
Additional Information

Number of data points 22
Proteins Fibronectin ; Fibronectin
Unique complexes 1
Assays/Quantities/Protocols Experimental Assay: Tm pH:7.0, buffers:Sodium phosphate: 50 mM, prot_conc:4 micro M ; Experimental Assay: Cm pH:7.0, buffers:Sodium phosphate: 50 mM, prot_conc:4 micro M ; Experimental Assay: m pH:7.0, buffers:Sodium phosphate: 50 mM, prot_conc:4 micro M ; Experimental Assay: dG_H2O pH:7.0, buffers:Sodium phosphate: 50 mM, prot_conc:4 micro M ; Experimental Assay: Cm prot_conc:1 micro M, buffers:Sodium phosphate: 50 mM, pH:7.0 ; Experimental Assay: m prot_conc:1 micro M, buffers:Sodium phosphate: 50 mM, pH:7.0 ; Experimental Assay: dG_H2O prot_conc:1 micro M, buffers:Sodium phosphate: 50 mM, pH:7.0 ; Experimental Assay: Cm pH:7.0, buffers:Sodium phosphate: 50 mM, prot_conc:25 micro M ; Experimental Assay: m pH:7.0, buffers:Sodium phosphate: 50 mM, prot_conc:25 micro M ; Experimental Assay: dG_H2O pH:7.0, buffers:Sodium phosphate: 50 mM, prot_conc:25 micro M ; Experimental Assay: Tm pH:5.0, prot_conc:4 micro MM, buffers:Sodium acetate: 50 mM ; Experimental Assay: Tm prot_conc:1 micro M, pH:5.0, buffers:Sodium acetate: 50 mM ; Experimental Assay: Cm prot_conc:0.12 mg/ml, pH:5.0, buffers:Sodium acetate: 50 mM ; Experimental Assay: m prot_conc:0.12 mg/ml, pH:5.0, buffers:Sodium acetate: 50 mM ; Experimental Assay: dG_H2O prot_conc:0.12 mg/ml, pH:5.0, buffers:Sodium acetate: 50 mM
Libraries Mutations for sequence VSDVPRDLEVVAATPTSLLISWDAPAVTVRYYRITYGETGGNSPVQEFTVPGSKSTATISGLKPGVDYTITVYAVTGRGDSPASSKPISINYRT
Sequence Assay Result Units