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Article: Identification, isolation, and characterization of a 41-kilodalton protein from rat germ cell-conditioned medium exhibiting concentration- dependent dual biological activities

TitleIdentification, isolation, and characterization of a 41-kilodalton protein from rat germ cell-conditioned medium exhibiting concentration- dependent dual biological activities
Authors
Issue Date1997
PublisherThe Endocrine Society. The Journal's web site is located at http://endo.endojournals.org
Citation
Endocrinology, 1997, v. 138 n. 8, p. 3259-3268 How to Cite?
AbstractIn this report, we describe the purification of a novel protease with dual biological actions from germ cell-conditioned medium (GCCM) where germ cells were isolated from adult rat testes using a mechanical procedure. Using multiple HPLC columns and two sequential high performance electrophoresis chromatography steps in association with an [125I]-collagen film assay to detect protease activity, a 41-kDa polypeptide (41-kDa-P) was purified to apparent homogeneity from GCCM. Partial N-terminal amino acid sequence analysis of the purified protein revealed a sequence of NH2-KYEFYEIXLL that, when compared with the existing database at Protein Identification Resource (PIR), GenBank, and BLAST revealed that this is a unique protein. The purified protein, when incubated with [125I]-testin, a Sertoli cell secretary product that is localized at the intertesticular cell junction and is resistant to tryptic digest, was found capable of hydrolyzing testin dose dependently. The proteolysis of [125I]-testin by this 41-kDa protein was inhibited by α2-macroglobulin (a Sertoli cell secretary product) also in a dose-dependent manner. A study on the interactions between different classes of protease inhibitors and the purified 41-kDa protein revealed that it is a serine protease. At doses ranging between 0.5 and 50 ng/ml, 41-kDa-P induced a dose-dependent inhibition of Sertoli cell secretory function using testin and clusterin as markers without any apparent proteolytic activity. However, at doses greater than 0.5 μg/ml, 41-kDa-P was found to cleave [125I]- collagen and [125I]-testin at physiological pH, indicating that this 41- kDa protein has dual biological activities whose primary action is concentration dependent. In view of the biological activities of this protease, it is postulated that this protein may be involved in facilitating germ cell migration in the epithelium.
Persistent Identifierhttp://hdl.handle.net/10722/49350
ISSN
2015 Impact Factor: 4.159
2015 SCImago Journal Rankings: 2.363
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorAravindan, GRen_HK
dc.contributor.authorMruk, Den_HK
dc.contributor.authorLee, WMen_HK
dc.contributor.authorCheng, CYen_HK
dc.date.accessioned2008-06-12T06:40:05Z-
dc.date.available2008-06-12T06:40:05Z-
dc.date.issued1997en_HK
dc.identifier.citationEndocrinology, 1997, v. 138 n. 8, p. 3259-3268en_HK
dc.identifier.issn0013-7227en_HK
dc.identifier.urihttp://hdl.handle.net/10722/49350-
dc.description.abstractIn this report, we describe the purification of a novel protease with dual biological actions from germ cell-conditioned medium (GCCM) where germ cells were isolated from adult rat testes using a mechanical procedure. Using multiple HPLC columns and two sequential high performance electrophoresis chromatography steps in association with an [125I]-collagen film assay to detect protease activity, a 41-kDa polypeptide (41-kDa-P) was purified to apparent homogeneity from GCCM. Partial N-terminal amino acid sequence analysis of the purified protein revealed a sequence of NH2-KYEFYEIXLL that, when compared with the existing database at Protein Identification Resource (PIR), GenBank, and BLAST revealed that this is a unique protein. The purified protein, when incubated with [125I]-testin, a Sertoli cell secretary product that is localized at the intertesticular cell junction and is resistant to tryptic digest, was found capable of hydrolyzing testin dose dependently. The proteolysis of [125I]-testin by this 41-kDa protein was inhibited by α2-macroglobulin (a Sertoli cell secretary product) also in a dose-dependent manner. A study on the interactions between different classes of protease inhibitors and the purified 41-kDa protein revealed that it is a serine protease. At doses ranging between 0.5 and 50 ng/ml, 41-kDa-P induced a dose-dependent inhibition of Sertoli cell secretory function using testin and clusterin as markers without any apparent proteolytic activity. However, at doses greater than 0.5 μg/ml, 41-kDa-P was found to cleave [125I]- collagen and [125I]-testin at physiological pH, indicating that this 41- kDa protein has dual biological activities whose primary action is concentration dependent. In view of the biological activities of this protease, it is postulated that this protein may be involved in facilitating germ cell migration in the epithelium.en_HK
dc.format.extent418 bytes-
dc.format.mimetypetext/html-
dc.languageengen_HK
dc.publisherThe Endocrine Society. The Journal's web site is located at http://endo.endojournals.orgen_HK
dc.relation.ispartofEndocrinologyen_HK
dc.rightsEndocrinology. Copyright © The Endocrine Society.en_HK
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.subject.meshCulture Media, Conditioned - chemistry - pharmacologyen_HK
dc.subject.meshMolecular Chaperonesen_HK
dc.subject.meshPeptides - analysis - chemistry - metabolismen_HK
dc.subject.meshSerine Endopeptidases - analysis - chemistry - metabolismen_HK
dc.subject.meshTestis - cytology - metabolismen_HK
dc.titleIdentification, isolation, and characterization of a 41-kilodalton protein from rat germ cell-conditioned medium exhibiting concentration- dependent dual biological activitiesen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0013-7227&volume=138&issue=8&spage=3259&epage=3268&date=1997&atitle=Identification,+isolation,+and+characterization+of+a+41-kilodalton+protein+from+rat+germ+cell-conditioned+medium+exhibiting+concentration-dependent+dual+biological+activitiesen_HK
dc.identifier.emailLee, WM: hrszlwm@hku.hken_HK
dc.identifier.authorityLee, WM=rp00728en_HK
dc.description.naturepublished_or_final_versionen_HK
dc.identifier.doi10.1210/en.138.8.3259en_HK
dc.identifier.pmid9231776en_HK
dc.identifier.scopuseid_2-s2.0-0030873333en_HK
dc.identifier.hkuros22390-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0030873333&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume138en_HK
dc.identifier.issue8en_HK
dc.identifier.spage3259en_HK
dc.identifier.epage3268en_HK
dc.identifier.isiWOS:A1997XL84200024-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridAravindan, GR=6602336336en_HK
dc.identifier.scopusauthoridMruk, D=6701823934en_HK
dc.identifier.scopusauthoridLee, WM=24799156600en_HK
dc.identifier.scopusauthoridCheng, CY=7404797787en_HK

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