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Five different adjuvants were used in combination with the PI\u2010PLC\u2010released proteins, and three of these proteins (24, 48, and 53 kDa) were shown to be potent antigens for immunization of female monkeys. The 53 kDa protein was found to be a surface coating protein and not a GPI\u2010anchored protein. Polyclonal antibodies to the 24 kDa protein and the 48 kDa protein were produced in rabbits. The two antibodies recognized both proteins on Western blots. The same rabbit antibodies recognized 28, 18, and 10 kDa bands on a Western blot of chemically reduced PI\u2010PLC\u2010released proteins, suggesting that the 48 kDa protein is a dimer of the 24 kDa protein, which we refer to as MAK248. Rabbit polyclonal antibodies developed to reduced fragments of the 24 kDa protein showed that the 18 and 10 kDa bands are proteolytic peptide fragments of the 24 kDa protein. Screening of tissues from male macaques showed that MAK248 is expressed only in the epididymis. Microsequencing of two proteolytic fragments of the 18 kDa component showed 100% amino acid homology to a 233 deduced amino acid sequence previously identified in human testes genome. Antibodies to MAK248 recognized a 24 kDa protein released from human sperm exposed to PI\u2010PLC. Antibodies to MAK248 recognized the equatorial segment and posterior head regions of capacitated cynomolgus macaque sperm. Structural analysis suggests that MAK248 is a novel CRISP protein and a member of the CAP (CRISP, Ag 5, PR\u20101) family of proteins. Based on amino acid sequence homology, it is possible that MAK248 functions as a protease inhibitor. Mol. Reprod. Dev. 63: 488\u2013499, 2002. \u00a9 2002 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/mrd.10193","type":"journal-article","created":{"date-parts":[[2002,11,4]],"date-time":"2002-11-04T13:55:39Z","timestamp":1036418139000},"page":"488-499","source":"Crossref","is-referenced-by-count":17,"title":["Identification of a novel GPI\u2010anchored CRISP glycoprotein, MAK248, located on the posterior head and equatorial segment of cynomolgus macaque 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