Calpastatin gene in human testis.
Antibodies present in a serum obtained from an infertile woman interacted with a 17.5 kD glycoprotein (BS-17 component) extracted from human sperm by Western blot. Polyclonal antibodies were raised against the BS-17 component and used to identify positive staining clones from a human testis lambda gt11 expression library. The cDNA encoding the BS-17 component was isolated and its nucleotide sequence determined. The BS-17 cDNA contained 758 nucleotides with an open reading frame of 558 nucleotides encoding a polypeptide consisting of 186 amino acids. The BS-17 cDNA showed 99.7% homology in 758 nucleotides overlap with the 3' terminus of the gene coding calpastatin and 99.5% identity in 186 amino acid overlap with the carboxyl terminus of calpastatin. The BS-17 component of human sperm corresponds to the carboxyl terminus of calpastatin. This conclusion is supported by the finding that the polyclonal antibodies also interacted with a 84 kD protein corresponding to the M(r) of calpastatin.
National Laboratory of Medical Molecular Biology, Beijing, China.
L F
LF
Wang
S G
SG
Wei
S Y
SY
Miao
Q Y
QY
Liu
S S
SS
Koide
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D015153:Blotting, Western; D002135:Calcium-Binding Proteins / Q000235:genetics*; D003001:Cloning, Molecular; D015853:Cysteine Proteinase Inhibitors / Q000235:genetics*; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D005260:Female; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D013094:Spermatozoa / Q000276:immunology / Q000378:metabolism*; D013737:Testis / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D002135:Calcium-Binding Proteins; D015853:Cysteine Proteinase Inhibitors; D018076:DNA, Complementary; D007106:Immune Sera; C033668:calpastatin