Effect of nucleotides, divalent cations and temperature on the tryptic susceptibility of myosin subfragment 1.
The kinetics of tryptic breakdown of the heavy chain of chymotryptic myosin subfragment 1 (S1) according to the following scheme (where the numbers represent approximate masses in kDa) are altered at 21 degrees C by divalent (Formula: see text) cations (Me2+) and by ATP, ADP, adenosine 5'-[beta, gamma-imino]triphosphate or PPi, with or without Me2+. ATP or its analogs slow step 2 and accelerate steps 3 and 4, while Me2+ accelerates step 2. ATP and its analogs decrease the amount of a transient 27-kDa peptide [Hozumi, T. & Muhlrad, A. (1981) Biochemistry 20, 2945-2950]. We have found direct evidence for the suggestion in this reference that the 27-kDa peptide is not an obligatory precursor of the 25-kDa fragment and that ATP or ADP suppresses the formation of the larger N-terminal fragment rather than accelerates its breakdown. Cross-linking of sulfhydryl groups located in the 20-kDa fragment leads to trapping of MgADP in the N-terminal 25-kDa peptide [Wells, J.A. & Yount, R.G. (1980) Biochemistry 19, 1711-1717]; this process affects the tryptic fragmentation of S1 similarly to, but less effectively than, nucleotides. Acts-S1 formation prevents the effect of ATP on fragmentation. At 37 degrees C S1 loses ATPase activity; tryptic digestion proceeds more rapidly and the 50-kDa and 25-kDa fragments are degraded to small peptides. Nucleotides protect against the effects of higher temperature by producing conformational changes not only in the 27-kDa N-terminal portion (containing the putative nucleotide binding site) of the heavy chain of S1 but also in the 50-kDa peptide.
D000205:Actomyosin / Q000032:analysis; D000818:Animals; D001665:Binding Sites; D002413:Cations, Divalent / Q000378:metabolism; D015879:Myosin Subfragments / Q000032:analysis; D009218:Myosins / Q000032:analysis* / Q000378:metabolism; D009711:Nucleotides / Q000378:metabolism; D010446:Peptide Fragments / Q000032:analysis* / Q000378:metabolism; D011487:Protein Conformation; D011817:Rabbits; D013329:Structure-Activity Relationship; D013696:Temperature; D014357:Trypsin
D002413:Cations, Divalent; D015879:Myosin Subfragments; D009711:Nucleotides; D010446:Peptide Fragments; D000205:Actomyosin; D014357:Trypsin; D009218:Myosins