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Effects of adenylyl imidodiphosphate, a nonhydrolyzable adenosine triphosphate analog, on reactivated and rigor wave sea urchin sperm

dc.contributor.authorPenningroth, Stephen M.
dc.contributor.authorWitman, George B.
dc.date2022-08-11T08:08:04.000
dc.date.accessioned2022-08-23T15:41:04Z
dc.date.available2022-08-23T15:41:04Z
dc.date.issued1978-12-01
dc.date.submitted2008-12-15
dc.identifier.citation<p>J Cell Biol. 1978 Dec;79(3):827-32.</p>
dc.identifier.issn0021-9525 (Print)
dc.identifier.doi10.1083/jcb.79.3.827
dc.identifier.pmid153347
dc.identifier.urihttp://hdl.handle.net/20.500.14038/26542
dc.description.abstractA nonhydrolyzable ATP analog, adenylyl imidodiphosphate (AMP-PNP), has been used to study the role of ATP binding in flagellar motility. Sea urchin sperm of Lytechinus pictus were demembranated, reactivated, and locked in "rigor waves" by a modification of the method of Gibbons and Gibbons (11). Rigor wave sperm relaxed within 2 min after addition of 4 micrometer ATP, and reactivated upon addition of 10-12 micrometer ATP. The beat frequency of the reactivated sperm varied with ATP concentration according to Michaelis-Menten kinetics ("Km" = 0.24 mM; "Vmax" = 44 Hz) and was competitively inhibited by AMP-PNP (Ki" approximately to 8.1 mM). Rigor wave sperm were completely relaxed (straightened) within 2 min by AMP-PNP at concentrations of 2-4 mM. The possibilities that relaxation in AMP-PNP was a result of ATP contamination, AMP-PNP hydrolysis, or lowering of the free Mg++ concentration were conclusively ruled out. The results suggest that dynein cross-bridge release is dependent upon ATP binding but not hydrolysis.
dc.language.isoen_US
dc.relation<p><a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=153347&dopt=Abstract">Link to Article in PubMed</a></p>
dc.relation.urlhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC2110271
dc.subjectAdenosine Triphosphate
dc.subjectAdenylyl Imidodiphosphate
dc.subjectAnimals
dc.subjectDynein ATPase
dc.subjectMale
dc.subjectMicrotubules
dc.subjectProtein Binding
dc.subjectSea Urchins
dc.subjectSperm Motility
dc.subjectSpermatozoa
dc.subjectAnimal Experimentation and Research
dc.subjectCell Biology
dc.subjectCells
dc.subjectHeterocyclic Compounds
dc.subjectInvestigative Techniques
dc.subjectNucleic Acids, Nucleotides, and Nucleosides
dc.subjectUrogenital System
dc.titleEffects of adenylyl imidodiphosphate, a nonhydrolyzable adenosine triphosphate analog, on reactivated and rigor wave sea urchin sperm
dc.typeJournal Article
dc.source.journaltitleThe Journal of cell biology
dc.source.volume79
dc.source.issue3
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/cellbiology_pp/46
dc.identifier.contextkey682199
html.description.abstract<p>A nonhydrolyzable ATP analog, adenylyl imidodiphosphate (AMP-PNP), has been used to study the role of ATP binding in flagellar motility. Sea urchin sperm of Lytechinus pictus were demembranated, reactivated, and locked in "rigor waves" by a modification of the method of Gibbons and Gibbons (11). Rigor wave sperm relaxed within 2 min after addition of 4 micrometer ATP, and reactivated upon addition of 10-12 micrometer ATP. The beat frequency of the reactivated sperm varied with ATP concentration according to Michaelis-Menten kinetics ("Km" = 0.24 mM; "Vmax" = 44 Hz) and was competitively inhibited by AMP-PNP (Ki" approximately to 8.1 mM). Rigor wave sperm were completely relaxed (straightened) within 2 min by AMP-PNP at concentrations of 2-4 mM. The possibilities that relaxation in AMP-PNP was a result of ATP contamination, AMP-PNP hydrolysis, or lowering of the free Mg++ concentration were conclusively ruled out. The results suggest that dynein cross-bridge release is dependent upon ATP binding but not hydrolysis.</p>
dc.identifier.submissionpathcellbiology_pp/46
dc.contributor.departmentDepartment of Cell Biology
dc.source.pages827-32


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