High Cu(I) and low proton affinities of the CXXC motif of Bacillus subtilis CopZ

2008 ◽  
Vol 413 (3) ◽  
pp. 459-465 ◽  
Author(s):  
Liang Zhou ◽  
Chloe Singleton ◽  
Nick E. Le Brun

CopZ, an Atx1-like copper chaperone from the bacterium Bacillus subtilis, functions as part of a complex cellular machinery for Cu(I) trafficking and detoxification, in which it interacts specifically with the transmembrane Cu(I)-transporter CopA. Here we demonstrate that the cysteine residues of the MXCXXC Cu(I)-binding motif of CopZ have low proton affinities, with both exhibiting pKa values of 6 or below. Chelator competition experiments demonstrated that the protein binds Cu(I) with extremely high affinity, with a small but significant pH-dependence over the range pH 6.5–8.0. From these data, a pH-corrected formation constant, β2=∼6×1022 M−2, was determined. Rapid exchange of Cu(I) between CopZ and the Cu(I)-chelator BCS (bathocuproine disulfonate) indicated that the mechanism of exchange does not involve simple dissociation of Cu(I) from CopZ (or BCS), but instead proceeds via the formation of a transient Cu(I)-mediated protein–chelator complex. Such a mechanism has similarities to the Cu(I)-exchange pathway that occurs between components of copper-trafficking pathways.

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Naomi Shimokawa-Chiba ◽  
Claudia Müller ◽  
Keigo Fujiwara ◽  
Bertrand Beckert ◽  
Koreaki Ito ◽  
...  

AbstractRescue of the ribosomes from dead-end translation complexes, such as those on truncated (non-stop) mRNA, is essential for the cell. Whereas bacteria use trans-translation for ribosome rescue, some Gram-negative species possess alternative and release factor (RF)-dependent rescue factors, which enable an RF to catalyze stop-codon-independent polypeptide release. We now discover that the Gram-positive Bacillus subtilis has an evolutionarily distinct ribosome rescue factor named BrfA. Genetic analysis shows that B. subtilis requires the function of either trans-translation or BrfA for growth, even in the absence of proteotoxic stresses. Biochemical and cryo-electron microscopy (cryo-EM) characterization demonstrates that BrfA binds to non-stop stalled ribosomes, recruits homologous RF2, but not RF1, and induces its transition into an open active conformation. Although BrfA is distinct from E. coli ArfA, they use convergent strategies in terms of mode of action and expression regulation, indicating that many bacteria may have evolved as yet unidentified ribosome rescue systems.


Heterocycles ◽  
2013 ◽  
Vol 87 (2) ◽  
pp. 307 ◽  
Author(s):  
Hee Jae Shin ◽  
Fakir Shahidullah Tareq ◽  
Ji Hye Kim ◽  
Min Ah Lee ◽  
Hyi-Seung Lee ◽  
...  

2015 ◽  
Vol 80 (12) ◽  
pp. 1589-1597 ◽  
Author(s):  
L. S. Khailova ◽  
P. A. Nazarov ◽  
N. V. Sumbatyan ◽  
G. A. Korshunova ◽  
T. I. Rokitskaya ◽  
...  

2004 ◽  
Vol 21 (2) ◽  
pp. 69-78 ◽  
Author(s):  
Patricia A. Maurice ◽  
Maciej Manecki ◽  
Jeremy B. Fein ◽  
Jennifer Schaefer

2021 ◽  
Vol 9 (1) ◽  
pp. 18-26
Author(s):  
Ade Maria Ulfa ◽  
◽  
Nofita Nofita ◽  
Bangun Saras Sandi ◽  
◽  
...  

ABSTRACT Sweat is produced by the apocrine glands, if infected by bacteria that play a role in the decay process will certainly produce foot odor. Some of the bacteria that cause, including Staphylococcus epidermis, Corynebacterium acne and there is one bacterium that causes pungent foot odor that is Bacillus subtilis. Bacillus subtilis enzyme leucine dehydrogenase produced the highest, resulting in isovaleric acid foot odor. Lime peel (Citrus aurantifolia) has the potential to be developed for the antibacterial active ingredient of foot odor contained in tannins, alkaloids and flavonoids. Spray can be effective for inhibition of feet due to water fleas or bacterial infections. The purpose of this research is to test the inhibitory zone of the preparation of foot odor spray ethanol extract of lime peel (Citrus aurantifolia) with variations of gelling agent. Bacterial inhibition zone testing on extracts of lime peel spray preparations using the disc method. This test was carried out on spray with extract concentration of 0% extract base carbopol, 0% extract base HPMC, 0.2% extract base carbopol, 0.2% extract base HPMC, 0.4% extract base carbopol, 0.4% extract base HPMC and positive control with an average inhibition zone of 9,13 mm, 9,12 mm, 11,86 mm, 11,29 mm, 13,17 mm, 12,30 mm, 8,13 mm against the bacterium Bacillus subtilis. Antibacterial test results were analyzed using ONE WAY ANOVA, the results of statistical analysis on the preparation of lime peel extract showed a significant inhibition zone difference of 0.000 (P = <0.05) between all concentrations. Lime peel extract spray is effective in inhibiting the bacterium Bacillus subtilis. Key words: Sweat of foot odor, Lime skin (Citrus aurantifolia), Spray, bacteri Bacillus subtilis


2019 ◽  
Author(s):  
Naomi Shimokawa-Chiba ◽  
Claudia Müller ◽  
Keigo Fujiwara ◽  
Bertrand Beckert ◽  
Koreaki Ito ◽  
...  

SummaryRescue of the ribosomes from dead-end translation complexes, such as those on truncated (non-stop) mRNA, is essential for the cell. Whereas bacteria use trans-translation for ribosome rescue, some Gram-negative species possess alternative and release factor (RF)-dependent rescue factors, which enable an RF to catalyze stop codon-independent polypeptide release. We now discover that the Gram-positive Bacillus subtilis has an evolutionarily distinct ribosome rescue factor named ResQ. Genetic analysis shows that B. subtilis requires the function of either trans-translation or ResQ for growth, even in the absence of proteotoxic stresses. Biochemical and cryo-EM characterization demonstrates that ResQ binds to non-stop stalled ribosomes, recruits homologous RF2, but not RF1, and induces its transition into an open active conformation. Although ResQ is distinct from E. coli ArfA, they use convergent strategies in terms of mode of action and expression regulation, indicating that many bacteria may have evolved as yet unidentified ribosome rescue systems.


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