TY  - JOUR
A1  - Reyes, Anibal M.
A1  - Vazquez, Diego S.
A1  - Zeida, Ari
A1  - Hugo, Martin
A1  - Dolores Pineyro, M.
A1  - Ines De Armas, Maria
A1  - Estrin, Dario
A1  - Radi, Rafael
A1  - Santos, Javier
A1  - Trujillo, Madia
T1  - PrxQ B from Mycobacterium tuberculosis is a monomeric, thioredoxin-dependent and highly efficient fatty acid hydroperoxide reductase
JF  - Free radical biology and medicine : the official journal of the Oxygen Society, a constituent member of the International Society for Free Radical Research
N2  - Mycobacterium tuberculosis (M. tuberculosis) is the intracellular bacterium responsible for tuberculosis disease (TD). Inside the phagosomes of activated macrophages, M. tuberculosis is exposed to cytotoxic hydroperoxides such as hydrogen peroxide, fatty acid hydroperoxides and peroxynitrite. Thus, the characterization of the bacterial antioxidant systems could facilitate novel drug developments. In this work, we characterized the product of the gene Rv1608c from M. tuberculosis, which according to sequence homology had been annotated as a putative peroxiredoxin of the peroxiredoxin Q subfamily (PrxQ B from M. tuberculosis or MtPrxQ B). The protein has been reported to be essential for M. tuberculosis growth in cholesterol-rich medium. We demonstrated the M. tuberculosis thioredoxin B/C-dependent peroxidase activity of MtPrxQ B, which acted as a two-cysteine peroxiredoxin that could function, although less efficiently, using a one-cysteine mechanism. Through steady-state and competition kinetic analysis, we proved that the net forward rate constant of MtPrxQ B reaction was 3 orders of magnitude faster for fatty acid hydroperoxides than for hydrogen peroxide (3x10(6) vs 6x10(3) M-1 s(-1), respectively), while the rate constant of peroxynitrite reduction was (0.6-1.4) x10(6) M-1 s(-1) at pH 7.4. The enzyme lacked activity towards cholesterol hydroperoxides solubilized in sodium deoxycholate. Both thioredoxin B and C rapidly reduced the oxidized form of MtPrxQ B, with rates constants of 0.5x10(6) and 1x10(6) M-1 s(-1), respectively. Our data indicated that MtPrxQ B is monomeric in solution both under reduced and oxidized states. In spite of the similar hydrodynamic behavior the reduced and oxidized forms of the protein showed important structural differences that were reflected in the protein circular dichroism spectra.
KW  - Mycobacterium tuberculosis
KW  - Peroxiredoxin
KW  - Thioredoxin
KW  - Peroxynitrite
KW  - Fatty acid hydroperoxides
KW  - Thiol-dependent peroxidase
KW  - Peroxidatic and resolving cysteine
Y1  - 2016
U6  - https://doi.org/10.1016/j.freeradbiomed.2016.10.005
SN  - 0891-5849
SN  - 1873-4596
VL  - 101
SP  - 249
EP  - 260
PB  - Elsevier
CY  - New York
ER  - 
TY  - GEN
A1  - Connor, Daniel Oliver
A1  - Zantow, Jonas
A1  - Hust, Michael
A1  - Bier, Frank Fabian
A1  - von Nickisch-Rosenegk, Markus
T1  - Identification of novel immunogenic proteins of Neisseria gonorrhoeae by phage display
T2  - Postprints der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe
N2  - Neisseria gonorrhoeae is one of the most prevalent sexually transmitted diseases worldwide with more than 100 million new infections per year. A lack of intense research over the last decades and increasing resistances to the recommended antibiotics call for a better understanding of gonococcal infection, fast diagnostics and therapeutic measures against N. gonorrhoeae. Therefore, the aim of this work was to identify novel immunogenic proteins as a first step to advance those unresolved problems. For the identification of immunogenic proteins, pHORF oligopeptide phage display libraries of the entire N. gonorrhoeae genome were constructed. Several immunogenic oligopeptides were identified using polyclonal rabbit antibodies against N. gonorrhoeae. Corresponding full-length proteins of the identified oligopeptides were expressed and their immunogenic character was verified by ELISA. The immunogenic character of six proteins was identified for the first time. Additional 13 proteins were verified as immunogenic proteins in N. gonorrhoeae.
T3  - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 541 
KW  - proteomic analysis
KW  - vaccine antigens
KW  - gene-expression
KW  - Mycobacterium tuberculosis
KW  - antimicrobial resistance
KW  - recombinant antibodies
KW  - Salmonella Thyphimurium
KW  - untreatable Gonorrhea
KW  - multidrug-resistant
KW  - Escherichia coli
Y1  - 2019
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-411077
SN  - 1866-8372
IS  - 541
ER  -