Receptor-mediated responses of sea urchin sperm membranes to speract or resact
The goal of this Dissertation was to isolate plasma membranes which would bind the peptides speract (Gly-Phe-Asp-Leu-Asn-Gly-Gly-Gly-Val-Gly) or resact (Cys-Val-Thr-Gly-Ala-Pro-Gly-Cys-Val-Gly-Gly-Gly-Arg-Leu-NH/sub 2/) and respond to receptor occupancy with biochemical changes similar to those observed in the intact cell. Plasma membrane-enriched vehicles were initially prepared from Lytechinus pictus spermatozoa. The membranes bound (/sup 125/I)-labeled speract; potencies for competition for binding of speract and analogs of speract were similar to those observed for the stimulation of respiration or elevation of cyclic nucleotide concentrations in the intact cell. A speract analog (GGG(/sup 125/I)(Y/sup 2/)-speract) was cross-linked to the receptor, and a single radiolabeled band was detected on Na dodecylSO/sub 2/ gels. Membranes were subsequently prepared from A. punctulata spermatozoa, and a resact analogue (/sup 125/i-(tyr/sup 1/,ser/sup 8/)resact) was shown to bind to the membranes. Resact, but not speract, competed with the radiolabeled ligand for binding. When membranes were prepared at pH 6.0 in the presence of fluoride, guanylate cyclase remained principally in an apparent 160,000 M/sub r/form; incubation of the membranes with (gamma-/sup 32/P)ATP resulted in the formation of /sup 32/P-labeled guanylate cyclase. The addition of resact to the membranes caused a shift in the apparent M/sub r/ (160,000 to 150,000), a loss of /sup 32/P, and a 70% reduction in guanylate cyclase activity. L. pictus spermatozoa were examined for similar responses.
- Research Organization:
- Vanderbilt Univ., Nashville, TN (USA)
- OSTI ID:
- 6701110
- Resource Relation:
- Other Information: Thesis (Ph. D.)
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
PEPTIDES
RADIORECEPTOR ASSAY
SPERMATOZOA
BIOCHEMISTRY
ATP
CELL MEMBRANES
ELECTROPHORESIS
IODINE 125
MOLECULAR WEIGHT
PHOSPHORUS 32
RECEPTORS
SEA URCHINS
ANIMALS
AQUATIC ORGANISMS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CELL CONSTITUENTS
CHEMISTRY
DAYS LIVING RADIOISOTOPES
ECHINODERMS
ELECTRON CAPTURE RADIOISOTOPES
GAMETES
GERM CELLS
INTERMEDIATE MASS NUCLEI
INVERTEBRATES
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
MEMBRANE PROTEINS
MEMBRANES
NUCLEI
NUCLEOTIDES
ODD-EVEN NUCLEI
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PHOSPHORUS ISOTOPES
PROTEINS
RADIOISOTOPES
TRACER TECHNIQUES
550601* - Medicine- Unsealed Radionuclides in Diagnostics
550201 - Biochemistry- Tracer Techniques