Light-induced proteolysis of myosin heavy chain by Rose Bengal-conjugated antibody complexes.
The specific light-induced, non-enzymatic digestion of chicken skeletal muscle myosin heavy chain by xanthene dye-conjugated antibodies is reported. The xanthene dye Rose Bengal was conjugated to either a mouse monoclonal anti-myosin primary specific antibody or to goat anti-mouse IgG secondary antibodies. Under our experimental conditions, visible light induced the non-enzymatic breakdown of myosin heavy chains when chicken skeletal muscle myosin either directly formed a complex with Rose Bengal-conjugated anti-myosin antibodies or indirectly formed a complex with anti-myosin antibody-Rose Bengal-conjugated secondary antibodies. The rate of the photochemical reaction depended on irradiation time and temperature. Although SDS-PAGE and immunoblot analyses showed that fragments migrating below the myosin heavy chain polypeptide predominated, these analyses also showed higher molecular mass polypeptides were generated.
Conlon
K A
KA
Department of Pharmacological Sciences, School of Medicine, University Hospital and Medical Center, State University of New York at Stony Brook, Stony Brook, NY 11794-8651, USA. conlon@pharm.sunysb.edu
Berrios
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D000918:Antibody Specificity; D002645:Chickens; D010450:Endopeptidases / Q000378:metabolism; D019085:Fluorescent Antibody Technique, Direct; D019084:Fluorescent Antibody Technique, Indirect; D005456:Fluorescent Dyes; D006041:Goats; D007074:Immunoglobulin G; D008027:Light; D051379:Mice; D018482:Muscle, Skeletal / Q000378:metabolism; D018995:Myosin Heavy Chains / Q000276:immunology / Q000378:metabolism*; D010777:Photochemistry; D012395:Rose Bengal; D013696:Temperature; D013997:Time Factors
D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; D007074:Immunoglobulin G; D012395:Rose Bengal; D010450:Endopeptidases; D018995:Myosin Heavy Chains