Evolutionarily conserved surface residues constitute actin binding sites of tropomyosin
Evolutionarily conserved surface residues constitute actin binding sites of tropomyosin
复制标题
DOI：
10.1073/pnas.1101221108
复制
发表时间：
2011-06-21
影响因子：
11.1
通讯作者：
Hitchcock-DeGregori, Sarah E.
中科院分区：
文献类型：
--
作者：
Barua, Bipasha;Pamula, Melissa C.;Hitchcock-DeGregori, Sarah E.
Tropomyosin (Tm) is a two-chained, alpha-helical coiled-coil protein that associates end-to-end to form a continuous strand along actin filaments and regulates the functions and stability of actin in eukaryotic muscle and nonmuscle cells. Mutations in Tm cause skeletal and cardiac myopathies. We applied a neoteric molecular evolution approach to gain insight into the fundamental unresolved question of what makes the Tm coiled coil an actin binding protein. We carried out a phylogenetic analysis of 70 coding sequences of Tm genes from 26 animal species, from cnidarians to chordates, and evaluated the substitution rates (omega) at individual codons to identify conserved sites. The most conserved residues at surface b, c, f heptad repeat positions were mutated in rat striated muscle alpha Tm and expressed in Escherichia coli. Each mutant had 3-4 sites mutated to Ala within the first half or the second half of periods 2-6. Actin affinity and thermodynamic stability were determined in vitro. Mutations in the first half of periods 2, 4, and 5 resulted in the largest reduction in actin affinity (>4-fold), indicating these mutations include residues in actin-binding sites. Mutations in the second half of the periods had a
