Expression and characterization of the human YWK-II gene, encoding a sperm membrane protein related to the alzheimer betaA4-amyloid precursorprotein.
The YWK-II cDNA, RSD-2, encoding a sperm membrane protein was isolated from a rat testis cDNA expression library. Using the RSD-2 insert in combination with rapid amplification of cDNA ends (RACE), the corresponding human gene was isolated from a human testis cDNA expression library. The human testis cDNA, HSD-2, is ... 3654 bp in length and contains an open reading frame of 763 codons. Hydropathicity analysis showed that the deduced polypeptide is a single strand transmembrane protein. The deduced polypeptide has partial homology with the amyloid precursor protein (APP) and high homology with the amyloid precursor homologue, APLP2/APPH. The YWK-II gene was mapped and assigned to human chromosome locus: 11q24-25. Northern blotting of various human tissue RNAs using the HSD-2 cDNA as a probe showed that the gene is transcribed ubiquitously. The cytoplasmic domain of HSD-2 was expressed in Escherichia coli. In-vitro studies showed that the recombinant polypeptide bound to a GTP-binding protein (G(o)) and was phosphorylated by protein kinase C and cdc2 kinase. In mammalian F11 cells, the recombinant polypeptide was found to be coupled to G(o). Thus, the YWK-II component has the characteristics of a G(o)-coupled receptor and may be involved in G(o)-mediated signal transduction pathway. Protein kinase C and cdc2 kinase may regulate this pathway in spermatozoa by phosphorylating the cytoplasmic domain of the YWK-II component.
Mesh Terms:
Alzheimer Disease, Amino Acid Sequence, Amyloid beta-Protein Precursor, Base Sequence, Binding Sites, Chromosome Mapping, Chromosomes, Human, Pair 11, Cytoplasm, DNA, Complementary, GTP-Binding Protein alpha Subunits, Gi-Go, GTP-Binding Proteins, Gene Expression, Glutathione Transferase, Guanosine Triphosphate, Heterotrimeric GTP-Binding Proteins, Humans, Male, Membrane Proteins, Molecular Sequence Data, Nerve Tissue Proteins, Phosphorylation, Recombinant Fusion Proteins, Spermatozoa, Sulfur Radioisotopes, Tissue Distribution
Alzheimer Disease, Amino Acid Sequence, Amyloid beta-Protein Precursor, Base Sequence, Binding Sites, Chromosome Mapping, Chromosomes, Human, Pair 11, Cytoplasm, DNA, Complementary, GTP-Binding Protein alpha Subunits, Gi-Go, GTP-Binding Proteins, Gene Expression, Glutathione Transferase, Guanosine Triphosphate, Heterotrimeric GTP-Binding Proteins, Humans, Male, Membrane Proteins, Molecular Sequence Data, Nerve Tissue Proteins, Phosphorylation, Recombinant Fusion Proteins, Spermatozoa, Sulfur Radioisotopes, Tissue Distribution
Mol Hum Reprod
Date: Dec. 01, 2000
PubMed ID: 11101689
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