Sperm mitochondria-associated cysteine-rich protein (SMCP) is an autoantigen in Lewis rats

Citation
Jc. Herr et al., Sperm mitochondria-associated cysteine-rich protein (SMCP) is an autoantigen in Lewis rats, BIOL REPROD, 61(2), 1999, pp. 428-435
Citations number
74
Language
INGLESE
art.tipo
Article
Categorie Soggetti
da verificare
Journal title
BIOLOGY OF REPRODUCTION
ISSN journal
0006-3363 → ACNP
Volume
61
Issue
2
Year of publication
1999
Pages
428 - 435
Database
ISI
SICI code
0006-3363(199908)61:2<428:SMCP(I>2.0.ZU;2-N
Abstract
A common repertoire of rat sperm antigens have previously been identified b y Western blotting of sperm proteins with sera obtained after vasectomy or isoimmunization with sperm. Aside from a determination of their apparent ma sses, however, the biochemical characteristics of these antigens have remai ned unknown. In this study, a rat testis cDNA expression library was screen ed with polyclonal antibodies obtained from rats immunized with isologous s permatozoa to identify and sequence a full-length clone encoding rat sperm mitochondria-associated cysteine-rich protein (SMCP). The open reading fram e of SMCP was expressed in the pET22b vector, and recombinant SMCP (rec-SMC P) was purified. Sera from rats that had been vasectomized or hyperimmunize d with isologous sperm specifically recognized rec-SMCP whereas preimmune s era from these experimental groups did not react. Rabbit antiserum produced to rec-SMCP recognized rec-SMCP on Western blots and precisely immunolocal ized SMCP to the mid-piece of rat sperm. On Western blots against sperm ext racts, the rabbit antibody recognized a major protein band of approximately 22-25 kDa that co-migrated with bands of identical mass that were recogniz ed by sera from hyperimmune or vasectomized rats. These findings demonstrat e that SMCP is a sperm autoantigen, recognized following vasectomy, and an isoantigen, recognized by antibodies generated through isologous immunizati on with sperm.