Two-dimensional polyacrylamide gel electrophoresis has been used to map the pattern of protein synthesis in rat ventral prostate from both castrate and testosterone treated animals. Proteins were detected by fluorography of 35S-methionine incorporation. In the soluble or cytosol fraction, four major proteins that have molecular weights less than 25,000 and isoelectric points (pI) less then 6.0 and one than has a molecular weight above 30,000 and pI 6.6 are under androgen control. One of these proteins with molecular weight 12,000 and pI 5.4 also appears in the microsomal fraction. Using nonequilibrium pH gradient electrophoresis, soluble proteins of pI greater than 7.0 were investigated. One major protein of 20-25,000 daltons and pI 8.0 appears to be androgen dependent. The ventral prostate from castrate synthesized proteins which are not detectable in androgen stimulated castrates.

International Journal of Peptide and Protein ResearchVolume 16, Issue 2 p. 143-146 COMPLETE AMINO ACID SEQUENCE OF A MAJOR SECRETORY PROTEIN FROM RAT SEMINAL VESICLE YU-CHING E. PAN, YU-CHING E. PAN National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina, U.S.A.Search for more papers by this authorALLEN B. SILVERBERG, ALLEN B. SILVERBERG National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina, U.S.A.Search for more papers by this authorSTEPHEN E. HARRIS, STEPHEN E. HARRIS National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina, U.S.A.Search for more papers by this authorSTEVEN S. L. LI, Corresponding Author STEVEN S. L. LI National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina, U.S.A.National Institute of Environmental Health Science National Institutes of Health P.O. Box 12233 Research Triangle Park North Carolina 27709 U.S.A.Search for more papers by this author YU-CHING E. PAN, YU-CHING E. PAN National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina, U.S.A.Search for more papers by this authorALLEN B. SILVERBERG, ALLEN B. SILVERBERG National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina, U.S.A.Search for more papers by this authorSTEPHEN E. HARRIS, STEPHEN E. HARRIS National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina, U.S.A.Search for more papers by this authorSTEVEN S. L. LI, Corresponding Author STEVEN S. L. LI National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina, U.S.A.National Institute of Environmental Health Science National Institutes of Health P.O. Box 12233 Research Triangle Park North Carolina 27709 U.S.A.Search for more papers by this author First published: August 1980 https://doi.org/10.1111/j.1399-3011.1980.tb02947.xCitations: 15AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Citing Literature Volume16, Issue2August 1980Pages 143-146 RelatedInformation

The effect of spironolactone on five androgen-dependent proteins in the ventral prostate of the rat was investigated by two-dimensional gel electrophoresis. Spironolactone was given to intact male, castrated and androgen-stimulated castrated rats. It has been shown that spironolactone had no influence on the synthesis or accumulation of the androgen-dependent proteins in intact animals. However, spironolactone suppressed the restoration of the major androgen-dependent protein of low molecular weight in castrated rats given testosterone. The mechanism by which spironolactone exerts its anti-androgenic activity was shown to be unrelated to its capacity to inhibit the synthesis or accumulation of the five androgen-dependent proteins studied in this investigation.

Total poly (A)-containing messenger RNA [poly(A)-mRNAT] was isolated from seminal vesicles of adult male rats. By preparative 5%-20% linear sucrose gradient centrifugation in 1% SDS, a single peak was seen in the 11S region [poly(A)-mRNA11S]. Poly (A)-mRNAT and poly(A)-mRNA11S were translated in the wheat germ S-30 in vitro translation system. Laemmli-SDS-polyacrylamide slab gel electrophoresis of translation products of poly(A)-mRNAT revealed three major protein bands at approximately 18,500, 15,000, and 14,000 daltons. In vitro translation of poly(A)-mRNA11S across the 11S peak revealed partial purification of the mRNAs for the three predominant proteins. The lower molecular weight part of the 11S peak enriched for the mRNAs coding for the 15,000 and 14,000 dalton polypeptides, and the higher molecular weight side of the peak enriched for the 18,500 dalton mRNA. The highly abundant class of mRNA from rat seminal vesicle can be purified with one sucrose gradient centrifugation.

Total poly(A(+))-RNA (poly(A(+))-RNA(tot)) was isolated from rat seminal vesicle and its size distribution determined by 70% formamide 5-25% sucrose density analysis. One major peak was resolved in the 10-13 S region and accounted for approximately 35% of the total poly(A(+))-RNA applied. Preparative 1% SDS, 5-20% linear sucrose density gradients also resolved a single major peak in the 11S region (poly(A(+))(11S). Analysis of poly(A(+))-RNA(tot) and poly(A(+))-RNA(11S) under denaturing conditions on 2% agarose gel electrophoresis demonstrated two major components in both poly(A(+))-RNA populations. Size estimations for these components are 620 and 540 NT respectively. (3)H-cDNA was made to both poly(A(+))-RNA(tot) and poly(A(+))-RNA(11S). Back-hybridization of poly(A(+))-RNA(tot) and poly(A(+))-RNA(11S) to their respective (3)H-cDNA revealed a highly abundant class representing 41% and 85% of the sequences in their respective (3)H-cDNA's. The highly abundant class corresponded to 3-5 sequences present in 30,000-50,000 copies/cell. Invitro translation of poly(A(+))-RNA(11S) resulted in two major polypeptides coded for by the 620 NT long and 540 NT long poly(A(+))-RNA respectively.Images