title: Molecular cloning in the marmoset shows that semenogelin is not the precursor of the TRH-like peptide pGlu-Glu-Pro amide.
abstract: Two peptides with similar structures to thyrotropin-releasing hormone (TRH), pGlu-Glu-Pro amide and pGlu-Phe-Pro amide, have been identified in human seminal fluid and it has been shown that one of these peptides, pGlu-Glu-Pro amide, has the ability to increase the capacitation of sperm cells, consistent with a role in fertility. In order to select a species in which there is a high degree of expression of the genes that code for 'TRH-like' peptides, we have determined the levels of these peptides in the prostate, pancreas and thyroid of a range of species including rat, rabbit, ox, marmoset, macaque and man. The peptides were extracted from the tissues and purified before determination by RIA with TRH antibody. In addition, trypsin digestion and TRH RIA was used to investigate the presence of N-extended forms. The highest concentrations of TRH-immunoreactive peptides were found in the tissues of the marmoset, Callithrix jacchus. Ion-exchange chromatography demonstrated that marmoset thyroid contained principally authentic TRH, the pancreas contained both TRH and TRH-like peptides while the prostate contained TRH-like peptides alone. Further purification by HPLC showed that the main TRH-immunoreactive peptide in marmoset prostate was pGlu-Glu-Pro amide and a second component was identified as pGlu-Phe-Pro amide. The results indicate that the biosynthesis of these peptides could be studied to advantage in the marmoset. The biosynthetic precursors of the TRH-like peptides have not been identified. To examine whether pGlu-Glu-Pro amide might originate from semenogelin, we determined the sequence of semenogelin in the marmoset. It exhibited a high degree of homology with human semenogelin-I, but in place of the Lys-Gln-Glu-Pro sequence that might give rise to pGlu-Glu-Pro amide, marmoset semenogelin possessed the sequence Ser-Gln-Asp-Gln which cannot serve as a precursor for a TRH-like peptide. Further evidence was obtained by Northern blot analysis of a range of marmoset tissues. The results showed that semenogelin is not present in marmoset prostate. It is concluded that pGlu-Glu-Pro amide originates from a precursor distinct from semenogelin, both in marmoset and in man.
authors.affiliation: Institute of Molecular Biology, Billrothstrasse 11, A-5020 Salzburg, Austria.
authors.forename: A E
authors.initials: AE
authors.lastname: Huber
authors.forename: H
authors.initials: H
authors.lastname: Fraser
authors.forename: J
authors.initials: J
authors.lastname: del Rio-Garcia
authors.forename: G
authors.initials: G
authors.lastname: Kreil
authors.forename: D G
authors.initials: DG
authors.lastname: Smyth
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002144:Callithrix; D003001:Cloning, Molecular; D012739:Gonadal Steroid Hormones / Q000737:chemistry*; D006801:Humans; D008297:Male; D008322:Mammals; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry; D011467:Prostate / Q000737:chemistry; D011498:Protein Precursors / Q000737:chemistry; D011761:Pyrrolidonecarboxylic Acid / Q000031:analogs & derivatives; D012333:RNA, Messenger / Q000378:metabolism; D012661:Semen / Q000737:chemistry; D029607:Seminal Vesicle Secretory Proteins; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013973:Thyrotropin-Releasing Hormone / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000737:chemistry; D014357:Trypsin / Q000378:metabolism
chemical_list: D012739:Gonadal Steroid Hormones; D009842:Oligopeptides; D011498:Protein Precursors; D012333:RNA, Messenger; D029607:Seminal Vesicle Secretory Proteins; C424482:seminal vesicle-specific antigen; C074033:pyroglutamyl-phenylalanyl-prolinamide; D013973:Thyrotropin-Releasing Hormone; C059697:pyroglutamyl-glutamyl-proline amide; D014357:Trypsin; D011761:Pyrrolidonecarboxylic Acid