title: Characterization of the ATPase active site in myosin subfragment-1 with the use of vanadate plus ADP as a reversible "affinity-labeling" reagent: evidence for heterogeneity in the active sites.
abstract: Our previous work showed that the active site heterogeneity in heavy meromyosin (HMM) becomes evident when highly reactive SH-groups in HMM are modified by thimerosal (Kawamura, Higuchi, Emoto, & Tawada (1985) J. Biochem. 97, 1583-1593). The heterogeneity was revealed by "affinity-labeling" analysis with vanadate plus ADP, which was developed in the previous paper. To see whether this heterogeneity is due to the head-head interaction or two different alkali light chains present in HMM, we carried out similar studies with myosin subfragment-1 (S1) and one of the isozymes, S1(A1), which contains only the alkali light chain 1, and obtained essentially the same results as those previously obtained with HMM. The S1 results are easily explained by the same hypothesis previously used for explaining the HMM results: SH-modified S1 or S1(A1) contains two kinds of active site in a 1:1 ratio with almost the same ATPase activity: one hydrolyzes ATP by a mechanism giving a protein Trp fluorescence enhancement, whereas the other hydrolyzes ATP by another mechanism giving no fluorescence enhancement.
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000345:Affinity Labels; D001665:Binding Sites; D007700:Kinetics; D008433:Mathematics; D008954:Models, Biological; D015879:Myosin Subfragments / Q000378:metabolism; D009218:Myosins / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013849:Thimerosal / Q000494:pharmacology; D014638:Vanadates; D014639:Vanadium / Q000378:metabolism*
chemical_list: D000345:Affinity Labels; D015879:Myosin Subfragments; D010446:Peptide Fragments; D014639:Vanadium; D013849:Thimerosal; D014638:Vanadates; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D009218:Myosins