Antibacterial and Antifungal Activities of Vasostatin-1, the N-terminal Fragment of Chromogranin A

doi:10.1074/jbc.275.15.10745

Antibacterial and Antifungal Activities of Vasostatin-1, the N-terminal Fragment of Chromogranin A*

From ^‡INSERM Unité 338, “Biologie de la Communication Cellulaire,” 5 Rue Blaise Pascal 67084 Strasbourg Cedex, France, ^§Department of Biological and Technological Research, San Raffaele Scientific Institute, 20132 Milan, Italy,^¶CNRS UPR 4301, Centre de Biophysique Moléculaire, Rue Charles Sadron, 45071 Orléans, France, and ^‖CNRS, UPR 9022, Institut de Biologie Moléculaire et Cellulaire, 15 Rue René Descartes, 67084 Strasbourg Cedex, France

Abstract

Vasostatin-1, the natural N-terminal 1–76 chromogranin A (CGA)-derived fragment in bovine sequence, has been purified from chromaffin secretory granules and identified by sequencing and matrix-assisted laser desorption time-of-flight mass spectrometry. This peptide, which displays antibacterial activity against Gram-positive bacteria at micromolar concentrations, is also able to kill a large variety of filamentous fungi and yeast cells in the 1–10 μm range. We have found that the C-terminal moiety of vasostatin-1 is essential for the antifungal activity, and shorter active peptides have been synthesized. In addition, from the comparison with the activity displayed by related peptides (human recombinant and rat synthetic fragments), we could determine that antibacterial and antifungal activities have different structural requirements. To assess for such activities in vivo, CGA and CGA-derived fragments were identified in secretory material released from human polymorphonuclear neutrophils upon stimulation. Vasostatin-1, which is stored in a large variety of cells (endocrine, neuroendocrine, and neurons) and which is liberated from stimulated chromaffin and immune cells upon stress, may represent a new component active in innate immunity.

Footnotes

↵* This work was funded by INSERM and supported by grants from Meiji Institute of Health Science (Odawara, Japan), the Direction des Recherches, Etudes et Techniques Grant DRET 96-099 (to D. A.), Université Louis-Pasteur Contrats Pluriformation 93-96 and 97-2000 (to D. A.), the Ligue Contre le Cancer (to M. H. M. B.), the Association Recherche et Partage (to K. L.), the Fondation pour la Recherche Médicale (to K. L.), and the Région Alsace (to Y. G.).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.
↵** To whom correspondence should be addressed: INSERM U-338, 5 Rue Blaise Pascal 67084 Strasbourg Cedex, France. Tel.: 33-3-88-45-66-09; Fax: 33-3-88-60-08-06; E-mail: metz@neurochem.u-strasbg.fr.
↵2 M. H. Metz-Boutigue, K. Lugardon, R. Raffner, P. Gadroy, and D. Aunis, manuscript in preparation.
↵3 F. Sato, N. Ishida, T. Hasegawa, and H. Mukoyama, accession number Q9W7AO, DDBJ/EMBL/GenBank^TMdatabases.
Abbreviations:

CG

chromogranin

CGA

chromogranin A

MALDI-TOF

matrix-assisted laser desorption time-of-flight

PAGE

polyacrylamide gel electrophoresis

PEA

proenkephalin-A

PMNs

polymorphonuclear neutrophils

PTH

phenylthiohydantoin

VS-1

human recombinant Ser-Thr-Ala-CGA_1–78

HPLC

high pressure liquid chromatography
- Received November 8, 1999.
- Revision received January 7, 2000.
The American Society for Biochemistry and Molecular Biology, Inc.