title: Expression of POTE protein in human testis detected by novel monoclonal antibodies.
abstract: The POTE gene family is composed of 13 highly homologous paralogs preferentially expressed in prostate, ovary, testis, and placenta. We produced 10 monoclonal antibodies (MAbs) against three representative POTE paralogs: POTE-21, POTE-2gammaC, and POTE-22. One reacted with all three paralogs, six MAbs reacted with POTE-2gammaC and POTE-22, and three MAbs were specific to POTE-21. Epitopes of all 10 MAbs were located in the cysteine-rich repeats (CRRs) motifs located at the N-terminus of each POTE paralog. Testing the reactivity of each MAb with 12 different CRRs revealed slight differences among the antigenic determinants, which accounts for differences in cross-reactivity. Using MAbs HP8 and PG5 we were able to detect a POTE-actin fusion protein in human testis by immunoprecipitation followed by Western blotting. By immunohistochemistry we demonstrated that the POTE protein is expressed in primary spermatocytes, implying a role in spermatogenesis.
authors.affiliation: Laboratory of Molecular Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892-4264, USA.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Ise
authors.forename: Sudipto
authors.initials: S
authors.lastname: Das
authors.forename: Satoshi
authors.initials: S
authors.lastname: Nagata
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Maeda
authors.forename: Yoomi
authors.initials: Y
authors.lastname: Lee
authors.forename: Masanori
authors.initials: M
authors.lastname: Onda
authors.forename: Miriam R
authors.initials: MR
authors.lastname: Anver
authors.forename: Tapan K
authors.initials: TK
authors.lastname: Bera
authors.forename: Ira
authors.initials: I
authors.lastname: Pastan
mesh_terms: D000818:Animals; D017487:Ankyrins / Q000276:immunology / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000276:immunology; D002478:Cells, Cultured; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D013090:Spermatocytes / Q000276:immunology / Q000378:metabolism*; D013737:Testis / Q000276:immunology / Q000378:metabolism*
chemical_list: D017487:Ankyrins; D000911:Antibodies, Monoclonal