Protein thiols in spermatozoa and epididymal fluid of rats.
Thiol (SH) oxidation to disulphides (SS) is thought to be involved in sperm chromatin condensation and tail structure stabilization, which occur during maturation of spermatozoa. Previously developed procedures, using the fluorescent labelling agent monobromobimane (mBBr), enabled us to study the thiol-disulphide status of spermatozoa. Electrophoretic separation of labelled sperm proteins from the caput and cauda regions showed that during maturation thiol oxidation occurs in many protein fractions from the tail and that the magnitude of oxidation differs between proteins. Among the protein bands, one major band (MPB), probably a dense fibre constituent, is quantitatively prominent. N-Ethylmaleimide (NEM) or mBBr alkylation (of intact spermatozoa) changes the mobility of the caput MPB, but not that of the cauda MPB. The results indicated that the altered mobility of MPB is mainly due to a change in its shape, possibly resulting from the alkylation of a few critical SH groups. Epididymal fluid proteins contain both SH and SS. The thiol and disulphide content of the various epididymal proteins appears similar, although some diminution in fluorescence is seen in epididymal fluid proteins from the cauda region as compared with those from the caput region. The prominent changes in thiol status occur in the spermatozoa.
Seligman
J
J
Department of Human Genetics, Sackler School of Medicine, Tel Aviv University, Ramat Aviv, Israel.
Shalgi
R
R
D000818:Animals; D001643:Bridged Bicyclo Compounds; D003720:Densitometry; D004220:Disulfides / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004822:Epididymis / Q000737:chemistry / Q000378:metabolism*; D005456:Fluorescent Dyes; D008297:Male; D008856:Microscopy, Fluorescence; D011506:Proteins / Q000737:chemistry*; D051381:Rats; D011919:Rats, Inbred Strains; D013079:Sperm Maturation / Q000502:physiology*; D013094:Spermatozoa / Q000737:chemistry / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000032:analysis / Q000378:metabolism*
D001643:Bridged Bicyclo Compounds; D004220:Disulfides; D005456:Fluorescent Dyes; D011506:Proteins; D013438:Sulfhydryl Compounds; C028381:monobromobimane