Proteins of human semen. I. Two-dimensional mapping of human seminal fluid.

Edwards, Jesse J.; Tollaksen, Sandra L.; Anderson, Norman G.

doi:10.1093/clinchem/27.8.1335

Cited by 94 publications

(31 citation statements)

References 24 publications

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“…The tentative numerical assignments are given in an attempt to be consistent with the previous report (41: #2 alpha-2-macroglobulin, #5 ceruloplasmin, #9 alpha-1-beta-glycoprotein, #10 lactoferin, #I1 transfemn, #12 albumin, #18 Gc-globulin, #20 immunoglobulin heavy chain, #24 prostatic acid phosphatase, #26 alpha-1-acid glycoprotein, #27 haptoglobin beta chain, #29 actin, #40 immunoglobulin J chain, #42 immunoglobulin light chain, #44 apo a-1 lipoprotein. PAP isoenzymes, occupying position #24 @I 6.0-7.0, MW 45,000-50,000), have been reported in the seminal plasma [24], in the human urine [25], and in the prostatic secretions [4]. The lower molecular weight groups of proteins that showed a positive cross-reactivity with the PAP antibody are likely to be the PAP degradative fragments in the prostatic secretions.…”

Section: Discussionmentioning

confidence: 98%

Two‐dimensional characterization of prostatic acid phosphatase, prostatic specific antigen and prostate binding protein in expressed prostatic fluid

et al. 1986

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Specimens of pooled prostatic fluid, collected by rectal massage from men under 50 years of age with no apparent prostatic disorders, were subjected to two-dimensional gel electrophoresis to study the composition of its proteins. In a preliminary study, a total of 57 major protein groups were detected. In the present study, we attempted to identify, in the two-dimensional gels, those that are related to prostate-associated proteins, i.e., prostatic acid phosphatase (PAP), prostatic specific antigen (PSA), and prostate binding protein (PBP). Individual proteins were recognized by the procedure of Western Blot using specific antisera with peroxidase-antiperoxidase as the staining reagent. Each protein spot in the two-dimensional gel was expressed, along the abscissa, by its isoelectric point (pI) and, along the ordinate, by the molecular weight (MW). PAP consisted of a train of more than ten protein spots that occupied an area in the gel from pI 7.0, MW 45,000 to pI 6.0, MW 50,000. Four protein spots with a MW of 34,000 and a pI range of 8.2-8.8 were identified as PSA. PBP was observed as having three protein spots that were located at pI 5.6-6.6 with a single MW of 15,000. For PAP and PSA, additional protein spots with lower MWs also stained positively with the specific antisera, suggestive of the presence of degradative products of these proteins. Following the removal of the serum-related proteins by an extensive absorption with anti-human serum antibody by affinity chromatography, the prostatic fluid contained 27 major groups of non-serum proteins. These non-serum proteins in the prostatic fluid included PAP, PSA, PBP, and their related smaller molecular species. These results indicate that the prostatic fluid contains PAP, PSA, PBP and that their presence and the patterns of their distribution in the two-dimensional gels should be considered as the characteristic property of the prostatic secretions.

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Section: Discussionmentioning

confidence: 98%

Two‐dimensional characterization of prostatic acid phosphatase, prostatic specific antigen and prostate binding protein in expressed prostatic fluid

et al. 1986

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“…Following collection, the semen samples were treated as previously described (Edwards et al, 1981;Pulkkinen et al, 1977), with minor modifications. Briefly, the samples were frozen prior to liquefaction and stored at -7O"C, until further analysis.…”

Section: Lncapxenograji Tumorsmentioning

confidence: 99%

Detection and characterization of the prostate‐specific membrane antigen (PSMA) in tissue extracts and body fluids

Wright

1995

Intl Journal of Cancer

327

229

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The prostate-specific membrane antigen (PSMA) glycoprotein is recognized by the monoclonal antibody (MAb) 7E11-C5.3 as a predominant 100 kDa and minor 180 kDa component in LNCaP cell line extracts and its expression has been shown by immunohistochemistry to be highly restricted to prostate epithelium. The aim of the present study was to utilize Western blot analysis to determine if PSMA could be detected in human tissue extracts and body fluids and if so, which molecular forms were present. PSMA was detected as 120 and 200 kDa bands in normal, benign and malignant prostate tissues and seminal plasma. Further analysis demonstrated that the larger molecular form of PSMA may be a dimer of the lower m.w. species. The PSMA glycoprotein was not detected in the majority of non-prostate tissue extracts examined except for a low yet significant amount in normal salivary gland, brain and small intestine, suggesting that PSMA may not be as prostate-specific as originally thought. Since the prostate-specific antigen (PSA) has been shown to be maximally shed into the serum in high-grade and metastatic prostate carcinomas, it was surprising that PSMA could not be detected in serum by Western blot analysis even in patients with actively progressive metastatic disease. Second generation antibodies generated against different epitopes may be required to determine if PSMA is shed into serum. Our results support the hypothesis that PSMA is a novel prostate biomarker.

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“…Mammalian seminal plasma (SP), a physiological secretion from multiple glands of the male reproductive tract, functions as a vehicle for ejaculated spermatozoa (Mann 1978) and is important for sperm function and survival (Edwards et al 1981). Several studies have provided direct evidence that specific components of SP, particularly proteins, are adsorbed onto the surface of ejaculated sperm (Fuller et al 1994;Watson 1995;Perez et al 1996;De Jonge 1999).…”

Section: Introductionmentioning

confidence: 99%

Identification of RSVP14 and RSVP20 Components by Two‐dimensional Electrophoresis and Western‐blotting

Cardozo

Fernández‐Juan

Cebrián‐Pérez

et al. 2007

Reprod Domestic Animals

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We have already shown that RSVP14 and RSVP20, two ram seminal plasma (SP) proteins postulated to be involved in sperm capacitation and gamete interaction can protect spermatozoa against cold-shock. In this study, we use two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) for the analysis of SP proteins of Rasa Aragonesa rams, using enhanced protein solubilization in the presence of tributyl phosphine (TBP) and a polyacrylamide linear gradient gel with a narrow pH range (4-7). The image analysis of the 2D map detected 195 protein spots, with isoelectric points (pIs) ranging from 4.5 to 6.6, and molecular weight (M(r)) from 11.7 to 90.4. Staining of 2D gels with Pro-Q Emerald 300 Glycoprotein Stain revealed that most significant proteins in ram SP are glycosylated. The removing of protein N-linked oligosaccharides improved the gel resolution. 2D-PAGE analysis of the whole fraction 6 (F6) separated from ram SP by exclusion chromatography showed six main protein spots, four (a, b, c, d) in the 14 kDa and two (e, f) in the 20 kDa region. Western-blot analyses indicated that the anti-P14 antibody recognized four spots on the SP map, 4, 5, 6 and 7, that matched with spots a, b, c, d of F6 map. The anti-P20 antibody recognized spots 13 and 14 of SP map that corresponded to spots e, f of F6 map. The deduced sequences by de novo sequencing evidenced that protein spots 7 and 13 have significant similarities to BSP family, while protein spots 4 and 14 did not appear to be homologous with any reported protein in the current mammalian Proteinbank databases.

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Proteins of human semen. I. Two-dimensional mapping of human seminal fluid.

Cited by 94 publications

References 24 publications

Two‐dimensional characterization of prostatic acid phosphatase, prostatic specific antigen and prostate binding protein in expressed prostatic fluid

Two‐dimensional characterization of prostatic acid phosphatase, prostatic specific antigen and prostate binding protein in expressed prostatic fluid

Detection and characterization of the prostate‐specific membrane antigen (PSMA) in tissue extracts and body fluids

Identification of RSVP14 and RSVP20 Components by Two‐dimensional Electrophoresis and Western‐blotting

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