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Infection and Immunity, June 2000, p. 3210-3218, Vol. 68, No. 6
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Brucella abortus and Its Closest Phylogenetic Relative, Ochrobactrum spp., Differ in Outer Membrane Permeability and Cationic Peptide Resistance

J. Velasco,1 J. A. Bengoechea,1,2 K. Brandenburg,2 B. Lindner,2 U. Seydel,2 D. González,1 U. Zähringer,2 E. Moreno,3 and I. Moriyón1,*

Departamento de Microbiología, Universidad de Navarra, Pamplona, Spain1; Forschungszentrum Borstel, Borstel, Germany2; and Escuela de Medicina Veterinaria, Universidad Nacional, Heredia, Costa Rica3

Received 20 December 1999/Returned for modification 8 February 2000/Accepted 9 March 2000

The outer membrane (OM) of the intracellular parasite Brucella abortus is permeable to hydrophobic probes and resistant to destabilization by polycationic peptides and EDTA. The significance of these unusual properties was investigated in a comparative study with the opportunistic pathogens of the genus Ochrobactrum, the closest known Brucella relative. Ochrobactrum spp. OMs were impermeable to hydrophobic probes and sensitive to polymyxin B but resistant to EDTA. These properties were traced to lipopolysaccharide (LPS) because (i) insertion of B. abortus LPS, but not of Escherichia coli LPS, into Ochrobactrum OM increased its permeability; (ii) permeability and polymyxin B binding measured with LPS aggregates paralleled the results with live bacteria; and (iii) the predicted intermediate results were obtained with B. abortus-Ochrobactrum anthropi and E. coli-O. anthropi LPS hybrid aggregates. Although Ochrobactrum was sensitive to polymyxin, self-promoted uptake and bacterial lysis occurred without OM morphological changes, suggesting an unusual OM structural rigidity. Ochrobactrum and B. abortus LPSs showed no differences in phosphate, qualitative fatty acid composition, or acyl chain fluidity. However, Ochrobactrum LPS, but not B. abortus LPS, contained galacturonic acid. B. abortus and Ochrobactrum smooth LPS aggregates had similar size and zeta potential (-12 to -15 mV). Upon saturation with polymyxin, zeta potential became positive (1 mV) for Ochrobactrum smooth LPS while remaining negative (-5 mV) for B. abortus smooth LPS, suggesting hindered access to inner targets. These results show that although Ochrobactrum and Brucella share a basic OM pattern, subtle modifications in LPS core cause markedly different OM properties, possibly reflecting the adaptive evolution of B. abortus to pathogenicity.

* Corresponding author. Mailing address: Departamento de Microbiología, Universidad de Navarra, Aptdo. 177, 31080 Pamplona, Spain. Phone: 34-948-425600. Fax: 34-948-425649. E-mail: imoriyón{at}unav.es.

Infection and Immunity, June 2000, p. 3210-3218, Vol. 68, No. 6
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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