title: The calcium ion dependence of scallop myosin ATPase activity.
abstract: The ATPase activity of scallop (Pecten maximus) striated adductor myosin and heavy meromyosin (HMM) have been investigated as a function of [Ca2+] using formycin triphosphate (FTP) as a fluorescent ATP analogue. The FTPase activity of the regulated fraction of these preparations was activated steeply over the range of 0.1 to 1 microM [Ca2+], implying the existence of a form of cooperativity that is intrinsic to the myosin heads. In addition to the previously characterised heterogeneity with respect to an unregulated fraction, the regulated fraction of HMM was resolved into two populations whose activities showed a slightly different dependency on [Ca2+]. This was revealed unambiguously at intermediate levels of activation where, in some experiments, the product release rate constants differed for the two populations by more than fivefold. At maximum relaxation or maximum activation, these rate constants differed by two- to three-fold and were not clearly resolved by the multiexponential fitting procedure. The populations might arise as a consequence of isoenzymes, modification during preparation or slowly interconverting conformers; Ca2+ binding itself being a rapid equilibrium process in both populations. FTP turnover by myosin could not be analysed in such detail because of the technical problems of measuring the fluorescence of a suspension of filaments, but the rates of the elementary steps appeared similar to those of HMM. The fraction of unregulated molecules in myosin preparations was comparable to that of HMM indicating that if it is a consequence of preparative damage, the modification must occur prior to tryptic digestion.
authors: Department of Biochemistry, University of Leicester, U.K.
authors: A R
authors: AR
authors: Walmsley
authors: G E
authors: GE
authors: Evans
authors: C R
authors: CR
authors: Bagshaw
mesh_terms: D000818:Animals; D002118:Calcium / Q000502:physiology*; D005453:Fluorescence; D005573:Formycins / Q000378:metabolism; D008974:Mollusca / Q000378:metabolism* / Q000502:physiology; D009132:Muscles / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D015879:Myosin Subfragments / Q000032:analysis / Q000378:metabolism / Q000502:physiology; D009218:Myosins / Q000032:analysis / Q000378:metabolism* / Q000502:physiology; D012265:Ribonucleotides / Q000378:metabolism; D013050:Spectrometry, Fluorescence
chemical_list: D005573:Formycins; D015879:Myosin Subfragments; D012265:Ribonucleotides; C032552:formycin triphosphate; D009218:Myosins; D002118:Calcium