The A31P missense mutation in cardiac myosin binding protein C alters protein structure but does not cause haploinsufficiency

Sabine J van Dijk; Kristina Bezold Kooiker; Stacy Mazzalupo; Yuanzhang Yang; Alla S Kostyukova; Debbie J Mustacich; Elaine R Hoye; Joshua A Stern; Mark D Kittleson; Samantha P Harris

doi:10.1016/j.abb.2016.01.006

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The A31P missense mutation in cardiac myosin binding protein C alters protein structure but does not cause haploinsufficiency

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The A31P missense mutation in cardiac myosin binding protein C alters protein structure but does not cause haploinsufficiency

Sabine J van Dijk, Kristina Bezold Kooiker, Stacy Mazzalupo, Yuanzhang Yang, Alla S Kostyukova, Debbie J Mustacich, Elaine R Hoye, Joshua A Stern, Mark D Kittleson and Samantha P Harris

Archives of biochemistry and biophysics, Vol.601, pp.133-140

07/01/2016

DOI: https://doi.org/10.1016/j.abb.2016.01.006

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https://hdl.handle.net/2376/109043

PMCID: PMC5843467

PMID: 26777460

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https://doi.org/10.1016/j.abb.2016.01.006View

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Abstract

Hypertrophic cardiomyopathy

Missense mutation

cMyBP-C

Animal models of cardiac disease

Mutations in MYBPC3, the gene encoding cardiac myosin binding protein C (cMyBP-C), are a major cause of hypertrophic cardiomyopathy (HCM). While most mutations encode premature stop codons, missense mutations causing single amino acid substitutions are also common. Here we investigated effects of a single proline for alanine substitution at amino acid 31 (A31P) in the C0 domain of cMyBP-C, which was identified as a natural cause of HCM in cats. Results using recombinant proteins showed that the mutation disrupted C0 structure, altered sensitivity to trypsin digestion, and reduced recognition by an antibody that preferentially recognizes N-terminal domains of cMyBP-C. Western blots detecting A31P cMyBP-C in myocardium of cats heterozygous for the mutation showed a reduced amount of A31P mutant protein relative to wild-type cMyBP-C, but the total amount of cMyBP-C was not different in myocardium from cats with or without the A31P mutation indicating altered rates of synthesis/degradation of A31P cMyBP-C. Also, the mutant A31P cMyBP-C was properly localized in cardiac sarcomeres. These results indicate that reduced protein expression (haploinsufficiency) cannot account for effects of the A31P cMyBP-C mutation and instead suggest that the A31P mutation causes HCM through a poison polypeptide mechanism that disrupts cMyBP-C or myocyte function.

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Title: The A31P missense mutation in cardiac myosin binding protein C alters protein structure but does not cause haploinsufficiency
Creators: Sabine J van Dijk - Department of Cellular and Molecular Medicine, University of Arizona, Tucson, AZ, USA
Kristina Bezold Kooiker - Department of Neurobiology, Physiology and Behavior, University of California, Davis, CA, USA
Stacy Mazzalupo - Department of Cellular and Molecular Medicine, University of Arizona, Tucson, AZ, USA
Yuanzhang Yang - Department of Cellular and Molecular Medicine, University of Arizona, Tucson, AZ, USA
Alla S Kostyukova - Voiland School of Chemical Engineering and Bioengineering, Washington State University, Pullman, WA, USA
Debbie J Mustacich - Department of Cellular and Molecular Medicine, University of Arizona, Tucson, AZ, USA
Elaine R Hoye - Department of Neurobiology, Physiology and Behavior, University of California, Davis, CA, USA
Joshua A Stern - Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California, Davis, CA, USA
Mark D Kittleson - Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California, Davis, CA, USA
Samantha P Harris - Department of Cellular and Molecular Medicine, University of Arizona, Tucson, AZ, USA
Publication Details: Archives of biochemistry and biophysics, Vol.601, pp.133-140
Academic Unit: Chemical Engineering and Bioengineering, School of
Publisher: Elsevier Inc
Identifiers: 99900547489401842
Language: English
Resource Type: Journal article