Primary structure and characterization of an androgen-stimulated autoantigen purified from mouse seminal-vesicle secretion.
A protein extract of mouse seminal-vesicle secretion was used to immunize mature mice (Balb/c) of both sexes. Results of Western-blot analyses for these secretory proteins indicated that only one minor protein component could be recognized by the autoantisera prepared from either autoimmunization of male mice or isoimmunization of female mice. The autoantigen was purified from seminal-vesicle secretion. The purified autoantigen retained the ability to induce autoantibody formation. The autoantigen has glycoprotein characteristics: the majority of the carbohydrate is N-linked and the remainder is O-linked. Rabbit antibodies to the autoantigen were used to isolate the corresponding cDNA from a mouse seminal-vesicle cDNA library. The primary structure deduced from the cDNA sequence was confirmed by direct amino acid sequence determination. The results indicate that the core protein consists of 131 amino acid residues. Analysis of the primary structure indicates that the autoantigen has two potential acceptor sites for the N-linked carbohydrate at Asn-12 and Asn-122, three potential phosphorylation sites for casein kinase II at Thr-55, Ser-68 and Thr-76, and three potential phosphorylation sites for protein kinase C at Thr-28, Thr-40 and Thr-124. The core protein and the carbohydrate portion together have a molecular mass of 19 kDa. Results from Western- and Northern-blot analyses for various tissues indicate that the seminal vesicle is the sole organ producing this autoantigen. Expression of this autoantigen gene was stimulated by testosterone.
Yu
L C
LC
Institute of Biochemical Science, College of Science, National Taiwan University, Taipei, Republic of China.
Chen
J L
JL
Tsai
W B
WB
Chen
Y H
YH
D000595:Amino Acid Sequence; D000818:Animals; D001324:Autoantigens / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D015153:Blotting, Western; D002850:Chromatography, Gel; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006023:Glycoproteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000378:metabolism; D029607:Seminal Vesicle Secretory Proteins; D012669:Seminal Vesicles / Q000737:chemistry* / Q000378:metabolism; D013739:Testosterone / Q000502:physiology*
D001324:Autoantigens; D018076:DNA, Complementary; D006023:Glycoproteins; D012333:RNA, Messenger; D029607:Seminal Vesicle Secretory Proteins; C085073:seminal vesicle autoantigen; D013739:Testosterone