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Volume 272, Number 17, Issue of April 25, 1997 pp. 11109-11113
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

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Studies on the Energetics of Proaerolysin Secretion across the Outer Membrane of Aeromonas Species
EVIDENCE FOR A REQUIREMENT FOR BOTH THE PROTONMOTIVE FORCE AND ATP

(Received for publication, January 8, 1997)

Lucienne Letellier , S. Peter Howard ^§ and J. Thomas Buckley ^¶

From the  Laboratoire des Biomembranes, URA CNRS 1116, Université Paris-Sud, Bâtiment 432, F-91405 Orsay, France, the ^§ Department of Biology, University of Regina, Regina, Saskatchewan S4S 0A2, Canada, and the ^¶ Department of Biochemistry and Microbiology, University of Victoria, Box 3055, Victoria, British Columbia V8W 3P6, Canada

Aeromonas spp. secrete the channel-forming protein proaerolysin across their inner and outer membranes in separate steps using the general secretion pathway. Here we show that treating A. hydrophila or A. salmonicida with the protonophore carbonyl cyanide m-chorophenyl hydrazone blocks the second step in transport, secretion across the outer membrane from the periplasm, under conditions where the ATP levels in the cell are no different than the levels in control, secreting cells. A threshold for was observed in the region of 120 mV, below which secretion by both species was inhibited. Treatment of cells with arsenate, which lowered ATP levels but did not affect , also reduced secretion from the periplasm, an indication that there is an ATP requirement for this step independent of the requirement for . Secretion across the outer membrane was also arrested by increasing the osmotic pressure of the medium, even though cellular ATP levels and were not affected. This may be due to disruption of some necessary association between the inner and outer membranes.

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