Dual effect of actin on the accessibility of myosin essential light chain A1 to papain cleavage.
The influence of various amounts of actin on the proteolytic susceptibility of myosin essential light chain (ELC) A1, the binding of isolated A1 light chain and the N-peptide spanning N-terminal sequence of A1 to actin is studied to obtain more information on the role of the N-terminus of A1 light chain in the myosin-actin interaction. Low ratios of actin to myosin (1:1) lead to stimulate cleavage, whereas higher ratios (4:1) lead to protection of A1. Exposure of A1 by actin is especially seen in heavy meromyosin (HMM) and monomeric myosin and this is probably related to the full saturation of actin protomers with myosin heads. The protecting action of actin on A1 cleavage is more pronounced in myosin filaments. Conditions favoring the saturation of myosin regulatory light chain (RLC) with calcium ions instead of magnesium ions promotes the protection of A1. Cross-linking of HMM and actin results in higher yields of A1-actin product at high actin to myosin heads ratios. Isolated A1 light chain is pelleted by actin. A synthetic peptide spanning the N-terminal sequence of A1 can be cross-linked to actin. It is postulated that the protective action of actin on A1 papain cleavage is caused by the binding of the A1 N-terminus to actin. Changes in the RLC phosphorylation level and magnesium-for-calcium exchange in RLC may affect the probability of this interaction.
Nieznańska
H
H
Department of Muscle Biochemistry, Nencki Institute of Experimental Biology, Polish Academy of Sciences, Warsaw, Poland.
Nieznański
K
K
Efimova
N
N
Kakol
I
I
Stepkowski
D
D
D000199:Actins / Q000494:pharmacology*; D002118:Calcium / Q000494:pharmacology; D003432:Cross-Linking Reagents; D016014:Linear Models; D008274:Magnesium / Q000494:pharmacology; D018994:Myosin Light Chains / Q000378:metabolism*; D009994:Osmolar Concentration; D010206:Papain / Q000378:metabolism*; D011485:Protein Binding
D000199:Actins; D003432:Cross-Linking Reagents; D018994:Myosin Light Chains; D010206:Papain; D008274:Magnesium; D002118:Calcium