Cloning of a Glucose Phosphate Isomerase/Neuroleukin-Like Sperm Antigen Involved in Sperm Agglutination

• Published in: Biology of reproduction 2000-04, Vol.62 (4), p.1016-1023
• Main Authors: YAKIREVICH, E, NAOT, Y
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Antibodies, Monoclonal - immunology; Antibodies, Monoclonal - pharmacology; Antibody Specificity; Antigens - chemistry; Antigens - physiology; Base Sequence; Biological and medical sciences; Blotting, Western; Cloning, Molecular; Cross Reactions; DNA, Complementary - analysis; DNA, Complementary - biosynthesis; Electrophoresis, Polyacrylamide Gel; Female; Fluorescent Antibody Technique; Fundamental and applied biological sciences. Psychology; Glucose-6-Phosphate Isomerase - chemical synthesis; Male; Mammalian male genital system; Mice; Mice, Inbred BALB C; Molecular Sequence Data; Morphology. Physiology; Recombinant Proteins - analysis; Recombinant Proteins - biosynthesis; Sperm Agglutination - physiology; Sperm Motility - drug effects; Spermatozoa - immunology; Vertebrates: reproduction
• ISSN: 0006-3363; 1529-7268
• DOI: 10.1095/biolreprod62.4.1016

The mouse monoclonal antibody (mAb) A36 produced by us and shown to induce extensive, “tangled” sperm agglutination was used to isolate cDNAs encoding its cognate antigen. Three overlapping cDNA clones specifically recognized by the mAb were isolated from a human testis cDNA expression library in λgt11. Sequencing of these cDNAs yielded the complete nucleotide sequence of a 3-kilobase cDNA that encodes the mAb-related polypeptide, designated sperm antigen-36 (SA-36), composed of 558 deduced amino acids. SA-36 cDNA contained a 5′ untranslated region of 234 nucleotides (nt), an open reading frame of 1674 nt, and a 3′ untranslated region of 1138 nt. SA-36 cDNA displayed > 99% homology to glucose phosphate isomerase (GPI)/neuroleukin (NLK) mRNA. This surprising homology was confirmed in Western blots demonstrating that mAb A36 reacted specifically with GPI obtained from rabbit muscle and from baker's yeast. Moreover, polyclonal, monospecific antibodies produced against β-galactosidase/SA-36-3 fusion protein stained human spermatozoa and caused intensive agglutination of these cells in a manner similar to that with the mAb. Taken together, the data presented here demonstrated that mAb A36 cognate sperm surface antigen, encoded by SA-36 cDNA, is a GPI/NLK-like protein involved in sperm agglutination.