title: α-Tropomyosin with a D175N or E180G mutation in only one chain differs from tropomyosin with mutations in both chains.
abstract: α-Tropomyosin (Tm) carrying hypertrophic cardiomyopathy mutation D175N or E180G was expressed in Escherichia coli. We have assembled dimers of two polypeptide chains in vitro that carry one (αα*) or two (α*α*) copies of the mutation. We found that the presence of the mutation has little effect on dimer assembly, thereby predicting that individuals heterozygous for the Tm mutations are likely to express both αα* and α*α* Tm. Depending on the expression level, the heterodimer may be the predominant form in individuals carrying the mutation. Thus, it is important to define differences in the properties of Tm molecules carrying one or two copies of the mutation. We examined the Tm homo- and heterodimer properties: actin affinity, thermal stability, calcium regulation of myosin subfragment 1 binding, and calcium regulation of myofibril force. We report that the properties of the heterodimer may be similar to those of the wild-type homodimer (actin affinity, thermal stability, D175N αα*), similar to those of the mutant homodimer (calcium sensitivity, D175N αα*), intermediate between the two (actin affinity, E180G αα*), or different from both (thermal stability, E180G αα*). Thus, the properties of the homodimer are not a completely reliable guide to the properties of the heterodimer.
authors.affiliation: School of Biosciences, University of Kent, Canterbury, Kent, United Kingdom.
authors.forename: Miro
authors.initials: M
authors.lastname: Janco
authors.forename: Athanasia
authors.initials: A
authors.lastname: Kalyva
authors.forename: Beatrice
authors.initials: B
authors.lastname: Scellini
authors.forename: Nicoletta
authors.initials: N
authors.lastname: Piroddi
authors.forename: Chiara
authors.initials: C
authors.lastname: Tesi
authors.forename: Corrado
authors.initials: C
authors.lastname: Poggesi
authors.forename: Michael A
authors.initials: MA
authors.lastname: Geeves
mesh_terms: D000595:Amino Acid Sequence; D002942:Circular Dichroism; D019281:Dimerization; D008969:Molecular Sequence Data; D009154:Mutation; D011489:Protein Denaturation; D014335:Tropomyosin / Q000737:chemistry / Q000235:genetics*
chemical_list: D014335:Tropomyosin