Mechanistic heterogeneity in contractile properties of α-tropomyosin (TPM1) mutants associated with inherited cardiomyopathies

Tejas M. Gupte, Farah Haque, Binnu Gangadharan, Margaret S. Sunitha, Souhrid Mukherjee, Swetha Anandhan, Deepa Selvi Rani, Namita Mukundan, Amruta Jambekar, Kumarasamy Thangaraj, Ramanathan Sowdhamini, Ruth F. Sommese, Suman Nag, James A. Spudich, John A. Mercer

Research output: Contribution to journalArticle

24 Scopus citations

Abstract

The most frequent known causes of primary cardiomyopathies are mutations in the genes encoding sarcomeric proteins. Among those are 30 single-residue mutations in TPM1, the gene encoding α-tropomyosin. We examined seven mutant tropomyosins, E62Q, D84N, I172T, L185R, S215L, D230N, and M281T, that were chosen based on their clinical severity and locations along the molecule. The goal of our study was to determine how the biochemical characteristics of each of these mutant proteins are altered, which in turn could provide a structural rationale for treatment of the cardiomyopathies they produce. Measurements of Ca2+ sensitivity of human β-cardiac myosin ATPase activity are consistent with the hypothesis that hypertrophic cardiomyopathies are hypersensitive to Ca2+ activation, and dilated cardiomyopathies are hyposensitive. We also report correlations between ATPase activity at maximum Ca2+concentrations and conformational changes in TnC measured using a fluorescent probe, which provide evidence that different substitutions perturb the structure of the regulatory complex in different ways. Moreover, we observed changes in protein stability and protein-protein interactions in these mutants. Our results suggest multiple mechanistic pathways to hypertrophic and dilated cardiomyopathies. Finally, we examined a computationally designed mutant, E181K, that is hypersensitive, confirming predictions derived from in silico structural analysis.

Original languageEnglish (US)
Pages (from-to)7003-7015
Number of pages13
JournalJournal of Biological Chemistry
Volume290
Issue number11
DOIs
StatePublished - Mar 13 2015

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    Gupte, T. M., Haque, F., Gangadharan, B., Sunitha, M. S., Mukherjee, S., Anandhan, S., Rani, D. S., Mukundan, N., Jambekar, A., Thangaraj, K., Sowdhamini, R., Sommese, R. F., Nag, S., Spudich, J. A., & Mercer, J. A. (2015). Mechanistic heterogeneity in contractile properties of α-tropomyosin (TPM1) mutants associated with inherited cardiomyopathies. Journal of Biological Chemistry, 290(11), 7003-7015. https://doi.org/10.1074/jbc.M114.596676