Isolation and molecular characterization of AKAP110, a novel, sperm-specific protein kinase A-anchoring protein.
Agents that increase intracellular cAMP are potent stimulators of sperm motility. Anchoring inhibitor peptides, designed to disrupt the interaction of the cAMP-dependent protein kinase A (PKA) with A kinase-anchoring proteins (AKAPs), are potent inhibitors of sperm motility. These data suggest that PKA anchoring is a key biochemical mechanism controlling motility. We now report the isolation, identification, cloning, and characterization of AKAP110, the predominant AKAP detected in sperm lysates. AKAP110 cDNA was isolated and sequenced from mouse, bovine, and human testis libraries. Using truncated mutants, the RII-binding domain was identified. Alignment of the RII-binding domain on AKAP110 to those from other AKAPs reveals that AKAPs contain eight functionally conserved positions within an amphipathic helix structure that are responsible for RII interaction. Northern analysis of eight different tissues detected AKAP110 only in the testis, and in situ hybridization analysis detected AKAP110 only in round spermatids, suggesting that AKAP110 is a protein found only in male germ cells. Sperm cells contain both RI, located primarily in the acrosomal region of the head, and RII, located exclusively in the tail, regulatory subunits of PKA. Immunocytochemical analysis detected AKAP110 in the acrosomal region of the sperm head and along the entire length of the principal piece. These data suggest that AKAP110 shares compartments with both RI and RII isoforms of PKA and may function as a regulator of both motility- and head-associated functions such as capacitation and the acrosome reaction.
Pubmed ID: 10319321 RIS Download
A Kinase Anchor Proteins | Adaptor Proteins, Signal Transducing | Amino Acid Sequence | Animals | Binding Sites | Carrier Proteins | Cattle | Cloning, Molecular | Cyclic AMP-Dependent Protein Kinases | Humans | In Situ Hybridization | Male | Mice | Molecular Sequence Data | Organ Specificity | Proteins | RNA, Messenger | Seminal Plasma Proteins | Sequence Analysis | Sequence Homology, Amino Acid | Sperm Motility | Spermatozoa | Subcellular Fractions | Testis