Identification and characterization of a bovine myosin light chain-1 fast polymorphism.
This study aimed to identify genes or gene products associated with high lean muscle mass in bovines that may serve as potential markers for selection. An animal with a genetic predisposition to high lean muscle mass, the Belgium Blue, was chosen as a model to compare with the Holstein Friesian, a model that does not. Two-dimensional polyacrylamide gel electrophoresis analysis was utilized to compare the exhibited skeletal muscle proteome between the two animal types at two stages of foetal development. A previously uncharacterized polymorphism of a high expression myofibrillar protein, myosin light chain 1 fast (MLC-1f), was observed. The characterization of this polymorphism revealed a two amino acid insertion in a part of the protein that has been implicated in modulating myosin S1 ATPase activity. Furthermore, this polymorphism was shown to be the product of two alleles that are codominant. Screening studies were carried out on selected herds and showed a very high frequency of one allele. Both isoforms of MLC-1f were produced by recombinant means and purified. The recombinant proteins were exchanged into purified myosin hexamers that were then subject to assays measuring ATP consumption. The sensitivity of the assay utilized could not reveal any significant difference in ATPase activity between hexamers containing one or the other isoform.
Clarke
A J
AJ
School of Biological Sciences, University of Auckland, New Zealand. a.clarke@auckland.ac.nz
Knight
C
C
Bass
J
J
Cooper
G J
GJ
D000199:Actins / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D004247:DNA; D017931:DNA Primers; D018076:DNA, Complementary; D015180:Electrophoresis, Gel, Two-Dimensional; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000033:anatomy & histology; D018994:Myosin Light Chains / Q000235:genetics* / Q000378:metabolism; D011110:Polymorphism, Genetic; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013045:Species Specificity
D000199:Actins; D017931:DNA Primers; D018076:DNA, Complementary; D018994:Myosin Light Chains; D011994:Recombinant Proteins; D004247:DNA; D000251:Adenosine Triphosphatases