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Identification of the 48‐kDa G11 protein from guinea pig testes as sperad

Name: Identification of the 48‐kDa G11 protein from guinea pig testes as sperad
Creator: Isurani Ilayperuma
Keywords: Male, Sperm-Ovum Interactions, 0301 basic medicine, 0303 health sciences, Recombinant Fusion Proteins, Blotting, Western, Guinea Pigs, Molecular Sequence Data, Membrane Proteins, Polymerase Chain Reaction

descriptionPublicationkeyboard_double_arrow_right Article 29 Oct 2002 English Publisher:WileyJournal:Journal of Experimental Zoology, volume 293, pages 617-623 (issn: 0022-104X, eissn: 1097-010X,

Authors: Isurani Ilayperuma;

doi: 10.1002/jez.10182

pmid: 12410611

Identification of the 48‐kDa G11 protein from guinea pig testes as sperad

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Abstract

AbstractA protein found specifically in the membrane of spermatozoa called G11 has been implicated in sperm–egg binding and fusion. This study describes purification and identification of the G11 antigen. The G11 protein was purified using anion exchange chromatography, immunoaffinity chromatography and preparative SDS‐PAGE and was subjected to amino acid microsequencing by tandem mass spectrometry. Internal amino acid sequence data derived from the 48‐kDa G11 protein revealed that G11 is the recently discovered guinea pig sperm protein, sperad. Sperad is a transmembrane protein present in the periacrosomal plasma membrane of guinea pig sperm. The cytoplasmic domain of sperad was amplified from a guinea pig testes cDNA expression library by polymerase chain reaction and cloned into a prokaryotic gene expression vector, pGEX‐2T. The recombinant glutathione S‐transferase fusion protein was immunoblotted with monoclonal antibody G11. The results obtained from this study confirmed the monoclonal antibody G11 epitope location on the cytoplasmic domain of sperad. J. Exp. Zool. 293:617–623, 2002. © 2002 Wiley‐Liss, Inc.

Related Organizations

University of Otago
New Zealand

Keywords

Male, Sperm-Ovum Interactions, Recombinant Fusion Proteins, Blotting, Western, Guinea Pigs, Molecular Sequence Data, Membrane Proteins, Polymerase Chain Reaction, Spermatozoa, Protein Structure, Tertiary, Molecular Weight, Organ Specificity, Sequence Analysis, Protein, Testis, Animals, Electrophoresis, Polyacrylamide Gel, Amino Acid Sequence

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