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Applied and Environmental Microbiology, February 1999, p. 489-498, Vol. 65, No. 2
Department of Microbiology, College of
Biological Sciences, University of Guelph, Guelph, Ontario, Canada
N1G 2W1
Received 18 September 1998/Accepted 13 November 1998
Pseudomonas aeruginosa PAO1 produces two chemically
distinct types of lipopolysaccharides (LPSs), termed A-band LPS and
B-band LPS. The A-band O-side chain is electroneutral at physiological pH, while the B-band O-side chain contains numerous negatively charged
sites due to the presence of uronic acid residues in the repeat unit
structure. Strain PAO1 (A+ B+) and three
isogenic LPS mutants (A+ B
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Effect of O-Side-Chain-Lipopolysaccharide Chemistry
on Metal Binding
, A
B+, and A
B
) were studied to
determine the contribution of the O-side-chain portion of LPS to metal
binding by the surfaces of gram-negative cells. Transmission electron
microscopy with energy-dispersive X-ray spectroscopy was used to locate
and analyze sites of metal deposition, while atomic absorption
spectrophotometry and inductively coupled plasma-mass spectrometry were
used to perform bulk quantitation of bound metal. The results indicated
that cells of all of the strains caused the precipitation of gold as
intracellular, elemental crystals with a d-spacing of 2.43 Å. This type of precipitation has not been reported previously for
gram-negative cells and suggests that in the organisms studied gold
binding is not a surface-mediated event. All four strains bound similar
amounts of copper (0.213 to 0.222 µmol/mg [dry weight] of cells) at
the cell surface, suggesting that the major surface metal-binding sites
reside in portions of the LPS which are common to all strains (perhaps
the phosphoryl groups in the core-lipid A region). However, significant
differences were observed in the abilities of strains dps89
(A
B+) and AK1401 (A+
B
) to bind iron and lanthanum, respectively. Strain dps89
caused the precipitation of iron (1.623 µmol/mg [dry weight] of
cells) as an amorphous mineral phase (possibly iron hydroxide) on the cell surface, while strain AK1401 nucleated precipitation of lanthanum (0.229 µmol/mg [dry weight] of cells) as apiculate,
surface-associated crystals. Neither iron nor lanthanum precipitates
were observed on the cells of other strains, which suggests that the
combination of A-band LPS and B-band LPS produced by a cell may result
in a cell surface which promotes the formation of metal-rich
precipitates. We therefore propose that the negatively charged sites
located in the O-side chains are not directly responsible for the
binding of metallic ions; however, the B-band LPS molecule as a whole may contribute to overall cell surface properties which favor the
precipitation of distinct metal-rich mineral phases.
*
Corresponding author. Mailing address: Department of
Microbiology, College of Biological Sciences, University of Guelph,
Guelph, ON, Canada N1G 2W1. Phone: (519) 824-4120, ext. 3366. Fax:
(519) 837-1802. E-mail: tjb{at}micro.uoguelph.ca.
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