Biophysical characterization of OprB, a glucose-inducible porin of Pseudomonas aeruginosa.
OprB, a glucose-inducible porin of P. aeruginosa, was characterized by black lipid bilayer analysis and circular dichroism spectroscopy. Black lipid bilayer analysis of OprB revealed a single-channel conductance of 25 pS, the presence of a glucose binding site with a Ks for glucose of 380 +/- 40 mM, and the formation of channels with a strong selection for anions. Analysis of P. aeruginosa OprB circular dichroism spectra revealed a high beta sheet content (40%) which is within the range of that determined for other porins. Values obtained from black lipid bilayer analysis were compared to those previously obtained for OprB of P. putida [Saravolac et al. (1991). J. Bacteriol. 173, 4970-4976] and indicated extensive similarities in the single-channel conductance and glucose-binding properties of these two porins. Immunological and amino terminal sequence analysis revealed a high degree of homology. Of the first 14 amino terminal residues, 12 were identical. A major difference between the two porins was found in their ion selectivity. Whereas P. aeruginosa OprB is anion selective, P. putida OprB and other carbohydrate selective porins are known to be cation selective.
Wylie
J L
JL
Department of Microbiology, University of Manitoba, Winnipeg, Canada.
Bernegger-Egli
C
C
O'Neil
J D
JD
Worobec
E A
EA
D000465:Algorithms; D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D002942:Circular Dichroism; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology; D008051:Lipid Bilayers; D008969:Molecular Sequence Data; D018272:Porins / Q000737:chemistry*; D011550:Pseudomonas aeruginosa / Q000187:drug effects / Q000378:metabolism*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D001426:Bacterial Proteins; D008051:Lipid Bilayers; C086247:OprB protein, Pseudomonas; D018272:Porins; D005947:Glucose