Cross-linking of contractile proteins from skeletal muscle by treatment with microbial transglutaminase.
The action on muscle proteins of microbial transglutaminase (MTGase), which catalyzes the formation of a "zero-length" covalent cross-link between glutamine and lysine residues in peptides, was studied in order to define a basis for future application of MTGase cross-linking to the study of muscle protein interaction. We examined the cross-linking of skeletal muscle myosin, myosin subfragments, actin, and myofibrils by treatment with MTGase and the possible side-effects of the cross-linking on the enzymic activity of myosin, and found that the rod portions of myosin in myosin filaments were quickly cross-linked to each other by the action of MTGase, but myosin subfragment 1 was not cross-linked to actin. The MgATPase activities at 0.5 M KCl of myosin, heavy meromyosin, subfragment 1, and subfragment 1-actin were not significantly affected by the MTGase reaction. A very small fraction of the head portion of heavy meromyosin was cross-linked to actin in their rigor complexes by MTGase, and the ATPase activity at 0.5 M KCl of the cross-linked heavy meromyosin-actin complexes was slightly enhanced.
Department of Molecular Biology, Graduate School of Medical Science, Kyushu University, Fukuoka.
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YP
Huang
K
K
Seguro
M
M
Motoki
K
K
Tawada
D000199:Actins / Q000737:chemistry; D000251:Adenosine Triphosphatases / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D009124:Muscle Proteins / Q000737:chemistry*; D009210:Myofibrils / Q000737:chemistry*; D015879:Myosin Subfragments / Q000737:chemistry; D009218:Myosins / Q000737:chemistry; D011503:Transglutaminases / Q000378:metabolism*
D000199:Actins; D003432:Cross-Linking Reagents; D009124:Muscle Proteins; D015879:Myosin Subfragments; D011503:Transglutaminases; D000251:Adenosine Triphosphatases; D009218:Myosins