PMID: 8955283Dec 1, 1996Paper

Recombinant Treponema pallidum rare outer membrane protein 1 (Tromp1) expressed in Escherichia coli has porin activity and surface antigenic exposure

Journal of Bacteriology
D R BlancoM A Lovett

Abstract

We recently reported the cloning and sequencing of the gene encoding a 31-kDa Treponema pallidum subsp. pallidum rare outer membrane porin protein, designated Tromp1 (D. R. Blanco, C. I. Champion, M. M. Exner, H. Erdjument-Bromage, R. E. W. Hancock, P. Tempst, J. N. Miller, and M. A. Lovett, J. Bacteriol. 177:3556-3562, 1995). Here, we report the stable expression of recombinant Tromp1 (rTromp1) in Escherichia coli. rTromp1 expressed without its signal peptide and containing a 22-residue N-terminal fusion resulted in high-level accumulation of a nonexported soluble protein that was purified to homogeneity by fast protein liquid chromatography (FPLC). Specific antiserum generated to the FPLC-purified rTromp1 fusion identified on immunoblots of T. pallidum the native 31-kDa Tromp1 protein and two higher-molecular-mass oligomeric forms of Tromp1 at 55 and 80 kDa. rTromp1 was also expressed with its native signal peptide by using an inducible T7 promoter. Under these conditions, rTromp1 fractionated predominantly with the E. coli soluble and outer membrane fractions, but not with the inner membrane fraction. rTromp1 isolated from the E. coli outer membrane and reconstituted into planar lipid bilayers showed porin activity based on ...Continue Reading

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Citations

Feb 3, 2006·Expert Review of Vaccines·Paul A Cullen, Caroline E Cameron
Jan 20, 2000·Infection and Immunity·A Centurion-LaraS A Lukehart
Jun 24, 2005·Infection and Immunity·Matthew McKevittTimothy Palzkill
Jan 19, 2006·Clinical Microbiology Reviews·Rebecca E Lafond, Sheila A Lukehart
Aug 23, 2005·Infection and Immunity·Cheryl I ChampionMichael A Lovett
Mar 7, 2006·Journal of Clinical Microbiology·Mary Beth BrinkmanTimothy Palzkill

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