Effects of adenylyl imidodiphosphate, a nonhydrolyzable adenosine triphosphate analog, on reactivated and rigor wave sea urchin sperm
UMass Chan Affiliations
Department of Cell BiologyDocument Type
Journal ArticlePublication Date
1978-12-01Keywords
Adenosine TriphosphateAdenylyl Imidodiphosphate
Animals
Dynein ATPase
Male
Microtubules
Protein Binding
Sea Urchins
Sperm Motility
Spermatozoa
Animal Experimentation and Research
Cell Biology
Cells
Heterocyclic Compounds
Investigative Techniques
Nucleic Acids, Nucleotides, and Nucleosides
Urogenital System
Metadata
Show full item recordAbstract
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.Source
J Cell Biol. 1978 Dec;79(3):827-32.
DOI
10.1083/jcb.79.3.827Permanent Link to this Item
http://hdl.handle.net/20.500.14038/26542PubMed ID
153347Related Resources
ae974a485f413a2113503eed53cd6c53
10.1083/jcb.79.3.827