Identification of Xenopus laevis sperm and egg envelope binding components on nitrocellulose membranes.
Interacting egg envelope and sperm surface components were identified for Xenopus laevis using blotting methods. Sperm were extracted with sodium dodecyl sulfate (SDS), the extracted proteins separated by gel electrophoresis and blotted, and the blots treated with 125I-labeled heat solubilized envelopes. The converse experiment was also performed where envelope components were separated by gel electrophoresis, blotted, and the blots treated with 125I-labeled sperm components. Blotted sperm components with apparent molecular weights of 14K, 19K, 25K, and 35K selectively bound the solubilized envelopes. All of the envelope binding components were found to be localized on the sperm surface by radioiodinating intact sperm using Iodo-Gen. The blotted egg envelope component with an apparent molecular weight of 37K selectively bound to solubilized sperm components, and this binding was due to the protein moiety of the glycoprotein. 125I-labeled heat solubilized envelopes from unfertilized and fertilized eggs showed the same pattern of binding to blotted sperm components. Selected sulfated carbohydrates (fucoidan, dextran sulfate, and heparin, but not chondroitin sulfate) inhibited fertilization and binding of 125I-labeled heat solubilized envelopes to blotted sperm extract. Thus, the binding of heat solubilized envelopes to electrophoretically separated and blotted sperm proteins may reflect cellular interactions.
Lindsay
L L
LL
Department of Biochemistry and Biophysics, University of California, Davis 95616.
Hedrick
J L
JL
D000818:Animals; D002241:Carbohydrates / Q000494:pharmacology; D002462:Cell Membrane / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005306:Fertilization / Q000187:drug effects; D008297:Male; D008565:Membrane Proteins / Q000032:analysis*; D010063:Ovum / Q000032:analysis* / Q000648:ultrastructure; D013084:Sperm-Ovum Interactions; D013094:Spermatozoa / Q000032:analysis*; D014982:Xenopus laevis / Q000502:physiology*
D002241:Carbohydrates; D008565:Membrane Proteins