A scanning calorimetric study of unfolding equilibria in homodimeric chicken gizzard tropomyosins.
Using both circular dichroism (CD) and differential scanning calorimetry (DSC), several laboratories find that the thermal unfolding transitions of alpha alpha and beta beta homodimeric coiled coils of rabbit tropomyosin are multistate and display an overall unfolding enthalpy of near 300 kcal (mol dimer)(-1). In contrast, an extant CD study of beta beta and gamma gamma species of chicken gizzard tropomyosin concludes that their unfolding transitions are simple two-state transitions, with much smaller overall enthalpies (98 kcal mol(-1) for beta beta and 162 kcal mol(-1) for gamma gamma). However, these smaller enthalpies have been questioned, because they imply a concentration dependence of the melting temperatures that is far larger than observed by CD. We report here DSC studies of the unfolding of both beta beta and gamma gamma chicken gizzard homodimers. The results show that these transitions are very similar to those in rabbit tropomyosins in that 1) the overall unfolding enthalpy is near 300 kcal mol(-1); 2) the overall delta C(rho) values are significantly positive; 3) the various transitions are multistate, requiring at least two and as many as four domains to fit the DSC data. DSC studies are also reported on these homodimeric species of chicken gizzard tropomyosin with a single interchain disulfide cross-link. These results are also generally similar to those for the correspondingly cross-linked rabbit tropomyosins.
O'Brien
R
R
Department of Chemistry, Yale University, New Haven, Connecticut 06520-8107, USA.
Sturtevant
J M
JM
Wrabl
J
J
Holtzer
M E
ME
Holtzer
A
A
D000818:Animals; D002152:Calorimetry, Differential Scanning / Q000379:methods; D002645:Chickens; D002942:Circular Dichroism; D003432:Cross-Linking Reagents; D019281:Dimerization; D005895:Gizzard, Avian; D007700:Kinetics; D046911:Macromolecular Substances; D009130:Muscle, Smooth; D010084:Oxidation-Reduction; D011489:Protein Denaturation; D017510:Protein Folding; D011817:Rabbits; D014335:Tropomyosin / Q000737:chemistry*
D003432:Cross-Linking Reagents; D046911:Macromolecular Substances; D014335:Tropomyosin