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Open Access Research article

The C313Y Piedmontese mutation decreases myostatin covalent dimerisation and stability

Carlene S Starck12 and Andrew J Sutherland-Smith1*

Author Affiliations

1 Institute of Molecular BioSciences, Massey University, Private Bag 11-222, Palmerston North 4442, New Zealand

2 The Hospital for Sick Children, 555 University Ave, Toronto ON, M5G 1X8, Canada

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BMC Research Notes 2011, 4:442  doi:10.1186/1756-0500-4-442

Published: 24 October 2011

Abstract

Background

Myostatin is a key negative regulator of muscle growth and development, whose activity has important implications for the treatment of muscle wastage disorders. Piedmontese cattle display a double-muscled phenotype associated with the expression of C313Y mutant myostatin. In vivo, C313Y myostatin is proteolytically processed, exported and circulated extracellularly but fails to correctly regulate muscle growth. The C313Y mutation removes the C313-containing disulphide bond, an integral part of the characteristic TGF-β cystine-knot structural motif.

Results

Here we present in vitro analysis of the structure and stability of the C313Y myostatin protein that reveals significantly decreased covalent dimerisation for C313Y myostatin accompanied by a loss of structural stability compared to wild type. The C313Y myostatin growth factor, processed from full length precursor protein, fails to inhibit C2C12 myoblast proliferation in contrast to wild type myostatin. Although structural modeling shows the substitution of tyrosine causes structural perturbation, biochemical analysis of additional disulphide mutants, C313A and C374A, indicates that an intact cystine-knot motif is a major determinant in myostatin growth factor stability and covalent dimerisation.

Conclusions

This research shows that the cystine-knot structure is important for myostatin dimerisation and stability, and that disruption of this structural motif perturbs myostatin signaling.