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The Neisseria gonorrhoeae Obg protein is an essential ribosome-associated GTPase and a potential drug target.

Zielke RA, Wierzbicki IH, Baarda BI, Sikora AE - BMC Microbiol. (2015)

Bottom Line: Serious complications associated with these infections are frequent and include pelvic inflammatory disease, ectopic pregnancy, and infertility.The cellular levels of Obg reach a maximum in the early logarithmic phase and remain constant throughout bacterial growth.Characterization of the GC Obg at the molecular and functional levels presented herein may facilitate the future targeting of this protein with small molecule inhibitors and the evaluation of identified lead compounds for bactericidal activity against GC and other drug-resistant bacteria.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutical Sciences, College of Pharmacy, Oregon State University, 433 Weniger Hall, 103 SW Memorial Pl, Corvallis, OR, 97330, USA. Ryszard.Zielke@oregonstate.edu.

ABSTRACT

Background: Neisseria gonorrhoeae (GC) is a Gram-negative pathogen that most commonly infects mucosal surfaces, causing sexually transmitted urethritis in men and endocervicitis in women. Serious complications associated with these infections are frequent and include pelvic inflammatory disease, ectopic pregnancy, and infertility. The incidence of gonorrhea cases remains high globally while antibiotic treatment options, the sole counter measures against gonorrhea, are declining due to the remarkable ability of GC to acquire resistance. Evaluating of potential drug targets is essential to provide opportunities for developing antimicrobials with new mechanisms of action. We propose the GC Obg protein, belonging to the Obg/CgtA GTPase subfamily, as a potential target for the development of therapeutic interventions against gonorrhea, and in this study perform its initial functional and biochemical characterization.

Results: We report that NGO1990 encodes Obg protein, which is an essential factor for GC viability, associates predominantly with the large 50S ribosomal subunit, and is stably expressed under conditions relevant to infection of the human host. The anti-Obg antisera cross-reacts with a panel of contemporary GC clinical isolates, demonstrating the ubiquitous nature of Obg. The cellular levels of Obg reach a maximum in the early logarithmic phase and remain constant throughout bacterial growth. The in vitro binding and hydrolysis of the fluorescent guanine nucleotide analogs mant-GTP and mant-GDP by recombinant wild type and T192AT193A mutated variants of Obg are also assessed.

Conclusions: Characterization of the GC Obg at the molecular and functional levels presented herein may facilitate the future targeting of this protein with small molecule inhibitors and the evaluation of identified lead compounds for bactericidal activity against GC and other drug-resistant bacteria.

No MeSH data available.


Related in: MedlinePlus

The polyclonal rabbit anti-ObgGC antibodies cross-react with cell lysates of 36 diverse GC strains. Samples of whole-cell lysates derived from various GC isolates (as indicated) harvested from GCB were matched by equivalent OD600 units and resolved in 4-20 % Tris-glycine precast gels. The proteins were transferred onto nitrocellulose membrane and probed with polyclonal rabbit anti-ObgGC antisera
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Fig7: The polyclonal rabbit anti-ObgGC antibodies cross-react with cell lysates of 36 diverse GC strains. Samples of whole-cell lysates derived from various GC isolates (as indicated) harvested from GCB were matched by equivalent OD600 units and resolved in 4-20 % Tris-glycine precast gels. The proteins were transferred onto nitrocellulose membrane and probed with polyclonal rabbit anti-ObgGC antisera

Mentions: Subsequently, to examine expression of ObgGC, a diversified panel of GC isolates was utilized. This panel included common laboratory strains MS11, F62, and 1291, as well as 32 strains isolated from different gonorrhea patients from distinct geographical areas and at different time points. The anti-ObgGC antibodies detected a band of the same size in all examined GC isolates, albeit the level of expression varied between some strains (Fig. 7).Fig. 7


The Neisseria gonorrhoeae Obg protein is an essential ribosome-associated GTPase and a potential drug target.

Zielke RA, Wierzbicki IH, Baarda BI, Sikora AE - BMC Microbiol. (2015)

The polyclonal rabbit anti-ObgGC antibodies cross-react with cell lysates of 36 diverse GC strains. Samples of whole-cell lysates derived from various GC isolates (as indicated) harvested from GCB were matched by equivalent OD600 units and resolved in 4-20 % Tris-glycine precast gels. The proteins were transferred onto nitrocellulose membrane and probed with polyclonal rabbit anti-ObgGC antisera
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4487204&req=5

Fig7: The polyclonal rabbit anti-ObgGC antibodies cross-react with cell lysates of 36 diverse GC strains. Samples of whole-cell lysates derived from various GC isolates (as indicated) harvested from GCB were matched by equivalent OD600 units and resolved in 4-20 % Tris-glycine precast gels. The proteins were transferred onto nitrocellulose membrane and probed with polyclonal rabbit anti-ObgGC antisera
Mentions: Subsequently, to examine expression of ObgGC, a diversified panel of GC isolates was utilized. This panel included common laboratory strains MS11, F62, and 1291, as well as 32 strains isolated from different gonorrhea patients from distinct geographical areas and at different time points. The anti-ObgGC antibodies detected a band of the same size in all examined GC isolates, albeit the level of expression varied between some strains (Fig. 7).Fig. 7

Bottom Line: Serious complications associated with these infections are frequent and include pelvic inflammatory disease, ectopic pregnancy, and infertility.The cellular levels of Obg reach a maximum in the early logarithmic phase and remain constant throughout bacterial growth.Characterization of the GC Obg at the molecular and functional levels presented herein may facilitate the future targeting of this protein with small molecule inhibitors and the evaluation of identified lead compounds for bactericidal activity against GC and other drug-resistant bacteria.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutical Sciences, College of Pharmacy, Oregon State University, 433 Weniger Hall, 103 SW Memorial Pl, Corvallis, OR, 97330, USA. Ryszard.Zielke@oregonstate.edu.

ABSTRACT

Background: Neisseria gonorrhoeae (GC) is a Gram-negative pathogen that most commonly infects mucosal surfaces, causing sexually transmitted urethritis in men and endocervicitis in women. Serious complications associated with these infections are frequent and include pelvic inflammatory disease, ectopic pregnancy, and infertility. The incidence of gonorrhea cases remains high globally while antibiotic treatment options, the sole counter measures against gonorrhea, are declining due to the remarkable ability of GC to acquire resistance. Evaluating of potential drug targets is essential to provide opportunities for developing antimicrobials with new mechanisms of action. We propose the GC Obg protein, belonging to the Obg/CgtA GTPase subfamily, as a potential target for the development of therapeutic interventions against gonorrhea, and in this study perform its initial functional and biochemical characterization.

Results: We report that NGO1990 encodes Obg protein, which is an essential factor for GC viability, associates predominantly with the large 50S ribosomal subunit, and is stably expressed under conditions relevant to infection of the human host. The anti-Obg antisera cross-reacts with a panel of contemporary GC clinical isolates, demonstrating the ubiquitous nature of Obg. The cellular levels of Obg reach a maximum in the early logarithmic phase and remain constant throughout bacterial growth. The in vitro binding and hydrolysis of the fluorescent guanine nucleotide analogs mant-GTP and mant-GDP by recombinant wild type and T192AT193A mutated variants of Obg are also assessed.

Conclusions: Characterization of the GC Obg at the molecular and functional levels presented herein may facilitate the future targeting of this protein with small molecule inhibitors and the evaluation of identified lead compounds for bactericidal activity against GC and other drug-resistant bacteria.

No MeSH data available.


Related in: MedlinePlus