Vanadate-sensitized cleavage of dynein heavy chains by 365-nm irradiation of demembranated sperm flagella and its effect on the flagellar motility

Journal Article · Mon Jun 15 00:00:00 EDT 1987 · J. Biol. Chem.; (United States)

OSTI ID:6070983

Gibbons, B H; Gibbons, I R

Irradiation of demembranated flagella of sea urchin sperm at 365 nm in the presence of 0.05-1 mM MgATP and 5-10 microM vanadate (Vi) cleaves the alpha and beta heavy chains of the outer arm dynein at the same site and at about the same rate as reported previously for the solubilized dynein. The decrease in intact alpha and beta heavy chain material is biphasic, with about 80% being lost with a half-time of 8-10 min, and the remainder more slowly. Five other axonemal polypeptides of Mr greater than 350,000 are lost similarly, concomitant with the appearance of at least 9 new peptides of Mr 150,000-250,000. The motility of irradiated sperm flagella upon subsequent dilution into reactivation medium containing 1 mM ATP and 2.5 mM catechol shows a progressive decrease in flagellar beat frequency for irradiation times that produce up to about 50% cleavage of the dynein heavy chains; more prolonged irradiation causes irreversible loss of motility. Competition between photocleaved and intact outer arm dynein for rebinding to dynein-depleted sperm flagella shows that cleavage has little effect upon the ability for rebinding, although the cleaved dynein partially inhibits subsequent motility. Substitution of MnATP for the MgATP in the irradiation medium prevents the loss of all of the axonemal polypeptides during irradiation for up to 60 min and also protects the potential for subsequent flagellar motility.

• Research Organization: Univ. of Hawaii, Honolulu
• OSTI ID: 6070983
• Journal Information: J. Biol. Chem.; (United States), Vol. 262:17
• Country of Publication: United States
• Language: English

Related Subjects

63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
POLYPEPTIDES
CLEAVAGE
SPERMATOZOA
BEHAVIOR
BIOLOGICAL RADIATION EFFECTS
ATP
BIOLOGICAL HALF-LIFE
MOLECULAR WEIGHT
SEA URCHINS
VANADIUM
ANIMALS
AQUATIC ORGANISMS
BIOLOGICAL EFFECTS
CRYSTAL STRUCTURE
ECHINODERMS
ELEMENTS
GAMETES
GERM CELLS
INVERTEBRATES
METALS
MICROSTRUCTURE
NUCLEOTIDES
ORGANIC COMPOUNDS
PEPTIDES
PROTEINS
RADIATION EFFECTS
TRANSITION ELEMENTS
560120* - Radiation Effects on Biochemicals
Cells
& Tissue Culture