A dense core vesicle protein is restricted to the cortex of granules in the exocrine atrial gland of Aplysia california.
We have generated a monoclonal antibody (mAb) 5E10 which recognizes an antigen localized to dense core vesicles (DCVs) in the atrial gland of Aplysia californica. mAb5E10 immunoprecipitates an abundant 57-kDa glycoprotein (atrial gland granule-specific antigen, AGSA) which is a soluble component of atrial gland DCVs. Electron microscopy reveals that AGSA immunoreactivity is restricted to the region between the dense core, which contains neuropeptide immunoreactivity, and the membrane of atrial gland DCVs. AGSA was purified by immunoaffinity chromatography, and the amino acid sequences of both N-terminal and internal cyanogen bromide fragments were determined. This information was used to isolate a 2.8-kilobase cDNA which encodes a 47-kDa protein. The predicted amino acid sequence contains the micro-sequenced peptides, an N-terminal hydrophobic signal sequence, and four N-linked glycosylation sites, but does not contain any significant homologies to database sequences. Northern blots and light level immunocytochemistry demonstrate that the AGSA gene is specifically expressed in the atrial gland. The identification of a protein localized to the cortex of DCVs suggests that this region has a specialized role in the function of these vesicles.
Sossin
W S
WS
Department of Biological Sciences, Stanford University, California 94305.
Kreiner
T
T
Barinaga
M
M
Schilling
J
J
Scheller
R H
RH
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001048:Aplysia / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D003594:Cytoplasmic Granules / Q000032:analysis* / Q000648:ultrastructure; D004247:DNA / Q000235:genetics; D005088:Exocrine Glands / Q000032:analysis*; D005796:Genes; D007150:Immunohistochemistry; D007447:Invertebrate Hormones / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D008970:Molecular Weight
D000911:Antibodies, Monoclonal; D007447:Invertebrate Hormones; C026880:egg-laying hormone, Mollusca; D004247:DNA