Stability of two β-tropomyosin isoforms: effects of mutation Arg91Gly
In order to comprehend the domain structure of two β-tropomyosin (β-Tm) isoforms (skeletal muscle and smooth muscle β-Tm) and the influence of the disease-causing mutation Arg91Gly on it, we studied the thermal unfolding of these tropomyosin species by means of differential scanning calorimetry (DSC). Our results show that the studied point mutation dramatically decreases thermal stability of a significant part of both β-Tm isoforms (about a half of the molecule) that unfolds as a cooperative unit (calorimetric domain). We have assigned this domain to the N-terminal part of the molecule combining, in the case of smooth muscle β-Tm, DSC studies with measurements of temperature dependence of pyrene excimer fluorescence, whose decrease reflects dissociation of two β-Tm chains in the region of pyrene-labeled Cys-36. Interestingly, the destabilizing effect of the mutation spreads along the coiled-coil reflecting the high extent of cooperativity within this part of the β-Tm molecule.
Keywordsβ-tropomyosin Thermal unfolding Protein stability Differential scanning calorimetry
This work was supported in part by the Russian Foundation for Basic Research (grant 09-04-00339).
- Kremneva E, Boussouf S, Nikolaeva O, Maytum R, Geeves MA, Levitsky DI (2004) Effects of two familial hypertrophic cardiomyopathy mutations in α-tropomyosin, Asp175Asn and Glu180Gly, on the thermal unfolding of actin-bound tropomyosin. Biophys J 87:3922–3933. doi: 10.1529/biophysj.104.048793 PubMedCrossRefGoogle Scholar