Folding and stability of a fibronectin type III domain of human tenascin.
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.
Clarke
J
J
Centre for Protein Engineering, MRC Centre, Hills Road, Cambridge, CB2 2QH, U.K.
Hamill
S J
SJ
Johnson
C M
CM
D003001:Cloning, Molecular; D004926:Escherichia coli; D005353:Fibronectins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006358:Hot Temperature; D006801:Humans; D007700:Kinetics; D011489:Protein Denaturation; D017510:Protein Folding; D019063:Tenascin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013816:Thermodynamics; D014508:Urea
D005353:Fibronectins; D019063:Tenascin; D014508:Urea