title: Helical order in tarantula thick filaments requires the "closed" conformation of the myosin head.
abstract: Myosin heads are helically ordered on the thick filament surface in relaxed muscle. In mammalian and avian filaments this helical arrangement is dependent on temperature and it has been suggested that helical order is related to ATP hydrolysis by the heads. To test this hypothesis, we have used electron microscopy and image analysis to study the ability and temperature dependence of analogs of ATP and ADP.Pi to induce helical order in tarantula thick filaments. ATP or analogs were added to rigor myofibrils or purified thick filaments at 22 degrees C and 4 degrees C and the samples negatively stained. The ADP.Pi analogs ADP.AlF4 and ADP.Vi, and the ATP analogs ADP.BeFx, AMPPNP and ATPgammaNH2, all induced helical order in tarantula thick filaments, independent of temperature. In the absence of nucleotide, or in the presence of ADP or the ATP analog, ATPgammaS, there was no helical ordering. According to crystallographic and tryptophan fluorescence studies, all of these analogs, except ATPgammaS and ADP, induce the "closed" conformation of the myosin head (in which the gamma phosphate pocket is closed). We suggest that helical order requires the closed conformation of the myosin head but is not dependent on the hydrolysis of ATP.
authors.affiliation: Department of Cell Biology, University of Massachusetts Medical School, 55 Lake Avenue North, Worcester, MA 01655, USA. maria.zoghbi@umassmed.edu
authors.forename: M E
authors.initials: ME
authors.lastname: Zoghbi
authors.forename: J L
authors.initials: JL
authors.lastname: Woodhead
authors.forename: R
authors.initials: R
authors.lastname: Craig
authors.forename: R
authors.initials: R
authors.lastname: Padrón
mesh_terms: D000244:Adenosine Diphosphate / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000737:chemistry; D017607:Aluminum Compounds / Q000737:chemistry; D000818:Animals; D005459:Fluorides / Q000737:chemistry; D008854:Microscopy, Electron; D009218:Myosins / Q000737:chemistry*; D011487:Protein Conformation; D013112:Spiders
chemical_list: D017607:Aluminum Compounds; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D009218:Myosins; D005459:Fluorides; C032311:aluminum fluoride