Post-translational phosphorylation of the slow/beta myosin heavy chain isoform in adult rabbit masseter muscle.
Four different phenotypes of slow muscle fibers, characterized by differential epitope expression in the slow/beta myosin heavy chain (MyHC) isoform, have been identified in adult rabbit masseter muscle. We investigated the role of post-translational phosphorylation in the expression of these four phenotypes. Serial cryostat sections were treated either with alkaline phosphatase to dephosphorylate proteins in the tissue, or with a brain kinase solution and ATP to phosphorylate them, and then stained, using four antibodies that bind specifically to the slow/beta MyHC isoform. In sections pre-treated with phosphatase, immunoreactivity to antibody A4.840 was abolished, but it could be restored by subsequent kinase/ATP treatment or ATP alone, indicating that the expression of its epitope requires phosphorylation. Phosphatase treatment resulted in an exposure of the epitope for antibody A4.951 in cells that normally bind this antibody only weakly or not at all, but since heat treatment alone produced similar effects, the role of phosphorylation in this enhancement is less certain. Immunoreactivity to antibodies S58 and BA-D5 were not influenced by phosphatase pre-treatment. Kinase/ATP treatment was only effective in changing antibody binding when tissues already had been phosphatase treated. We interpret these results to mean that sites of potential phosphorylation may already be occupied by O-linked glycosylation.
Pol-Rodriguez
M M
MM
Department of Cell Biology, Emory University School of Medicine, Atlanta, GA 30322, USA.
Schwartz
G A
GA
English
A W
AW
D000818:Animals; D030161:Avian Proteins; D008406:Masseter Muscle / Q000378:metabolism*; D018995:Myosin Heavy Chains / Q000378:metabolism*; D010641:Phenotype; D010766:Phosphorylation; D020033:Protein Isoforms / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D011817:Rabbits
D030161:Avian Proteins; D020033:Protein Isoforms; C434475:slow MyHC3 protein, Coturnix japonica; D018995:Myosin Heavy Chains