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Originally published In Press as doi:10.1074/jbc.M001974200 on May 12, 2000

J. Biol. Chem., Vol. 275, Issue 32, 24740-24751, August 11, 2000
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Cloning and Expression of Secretagogin, a Novel Neuroendocrine- and Pancreatic Islet of Langerhans-specific Ca2+-binding Protein*

Ludwig WagnerDagger §, Olena OliyarnykDagger , Wolfgang GartnerDagger , Peter NowotnyDagger , Marion Groeger||, Klaus Kaserer**, Werner WaldhäuslDagger , and Mark S. PasternackDagger Dagger

From the Dagger  Department of Medicine III, || Department of Dermatology, and ** Department of Clinical Pathology, University of Vienna, A-1090 Vienna, Austria and the Dagger Dagger  Infectious Disease Units, Pediatric and Medical Services, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02129

We have cloned a novel pancreatic beta cell and neuroendocrine cell-specific calcium-binding protein termed secretagogin. The cDNA obtained by immunoscreening a human pancreatic cDNA library using the recently described murine monoclonal antibody D24 contains an open reading frame of 828 base pairs. This codes for a cytoplasmic protein with six putative EF finger hand calcium-binding motifs. The gene could be localized to chromosome 6 by alignment with GenBank genomic sequence data. Northern blot analysis demonstrated abundant expression of this protein in the pancreas and to a lesser extent in the thyroid, adrenal medulla, and cortex. In addition it was expressed in scant quantity in the gastrointestinal tract (stomach, small intestine, and colon). Thyroid tissue expression of secretagogin was restricted to C-cells. Using a sandwich capture enzyme-linked immunosorbent assay with a detection limit of 6.5 pg/ml, considerable amounts of constitutively secreted protein could be measured in tissue culture supernatants of stably transfected RIN-5F and dog insulinoma (INS-H1) cell clones; however, in stably transfected Jurkat cells, the protein was only secreted upon CD3 stimulation. Functional analysis of transfected cell lines expressing secretagogin revealed an influence on calcium flux and cell proliferation. In RIN-5F cells, the antiproliferative effect is possibly due to secretagogin-triggered down-regulation of substance P transcription.

* This work was supported in part by Austrian Science Foundation Grant P11782-Med.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to GenBankTM/EBI Data Bank with the accession number(s) Y16752.

§ To whom correspondence should be addressed: Dept. of Medicine III, Div. of Clinical Endocrinology & Metabolism, University of Vienna, Währinger Gürtel 18-20, A-1090 Vienna, Austria. Fax: 43-1-40400/7790; E-mail: ludwig.wagner@akh-wien.ac.at.

Recipient of a scholarship from the Austrian Academy of Science.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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