Porin activity of the native and recombinant outer membrane protein Oms28 of Borrelia burgdorferi Journal Article
| Authors: | Skare, J. T.; Champion, C. I.; Mirzabekov, T. A.; Shang, E. S.; Blanco, D. R.; Erdjument-Bromage, H.; Tempst, P.; Kagan, B. L.; Miller, J. N.; Lovett, M. A. |
| Article Title: | Porin activity of the native and recombinant outer membrane protein Oms28 of Borrelia burgdorferi |
| Abstract: | The outer membrane-spanning (Oms) proteins of Borrelia burgdorferi have been visualized by freeze-fracture analysis but, until recently, not further characterized. We developed a method for the isolation of B. burgdorferi outer membrane vesicles and described porin activities with single-channel conductances of 0.6 and 12.6 nS in 1 M KCl. By using both nondenaturing isoelectric focusing gel electrophoresis and fast-performance liquid chromatography separation after detergent solubilization, we found that the 0.6-nS porin activity resided in a 28-kDa protein, designated Oms28. The oms28 gene was cloned, and its nucleotide sequence was determined. The deduced amino acid sequence of Oms28 predicted a 257-amino-acid precursor protein with a putative 24-amino-acid leader peptidase I signal sequence. Processed Oms28 yielded a mature protein with a predicted molecular mass of 25,363 Da. When overproduced in Escherichia coli, the Oms28 porin fractionated in part to the outer membrane. Sodium dodecyl sulfate- polyacrylamide gel-purified recombinant Oms28 from E. coli retained functional activity as demonstrated by an average single-channel conductance of 1.1 nS in the planar lipid bilayer assay. These findings confirmed that Oms28 is a B. burgdorferi porin, the first to be described. As such, it is of potential relevance to the pathogenesis of Lyme borreliosis and to the physiology of the spirochete. |
| Keywords: | controlled study; nonhuman; united states; polymerase chain reaction; protein localization; protein analysis; bacteria (microorganisms); europe; cloning, molecular; bacterial protein; bacterial proteins; amino acid sequence; molecular sequence data; nucleotide sequence; escherichia coli; peptide fragments; recombinant proteins; recombinant protein; base sequence; chromatography, high pressure liquid; dna primers; molecular weight; bacterium isolation; electrophoresis, polyacrylamide gel; potassium chloride; outer membrane protein; ion channels; priority journal; article; borrelia burgdorferi; porin; lyme disease; borrelia burgdorferi group; porins; borrelia; ion conductance; spirochaetales |
| Journal Title: | Journal of Bacteriology |
| Volume: | 178 |
| Issue: | 16 |
| ISSN: | 0021-9193 |
| Publisher: | American Society for Microbiology |
| Date Published: | 1996-08-01 |
| Start Page: | 4909 |
| End Page: | 4918 |
| Language: | English |
| PUBMED: | 8759855 |
| PROVIDER: | scopus |
| PMCID: | PMC178274 |
| DOI: | 10.1128/jb.178.16.4909-4918.1996 |
| DOI/URL: | |
| Notes: | Article -- Export Date: 22 November 2017 -- Source: Scopus |
