Abstract:
:Gram-negative bacteria acquire ferric siderophores through TonB-dependent outer membrane transporters (TBDT). By fluorescence spectroscopic hgh-throughput screening (FLHTS), we identified inhibitors of TonB-dependent ferric enterobactin (FeEnt) uptake through Escherichia coli FepA (EcoFepA). Among 165 inhibitors found in a primary screen of 17,441 compounds, we evaluated 20 in secondary tests: TonB-dependent ferric siderophore uptake and colicin killing and proton motive force-dependent lactose transport. Six of 20 primary hits inhibited TonB-dependent activity in all tests. Comparison of their effects on [59Fe]Ent and [14C]lactose accumulation suggested several as proton ionophores, but two chemicals, ebselen and ST0082990, are likely not proton ionophores and may inhibit TonB-ExbBD. The facility of FLHTS against E. coli led us to adapt it to Acinetobacter baumannii We identified its FepA ortholog (AbaFepA), deleted and cloned its structural gene, genetically engineered 8 Cys substitutions in its surface loops, labeled them with fluorescein, and made fluorescence spectroscopic observations of FeEnt uptake in A. baumannii Several Cys substitutions in AbaFepA (S279C, T562C, and S665C) were readily fluoresceinated and then suitable as sensors of FeEnt transport. As in E. coli, the test monitored TonB-dependent FeEnt uptake by AbaFepA. In microtiter format with A. baumannii, FLHTS produced Z' factors 0.6 to 0.8. These data validated the FLHTS strategy against even distantly related Gram-negative bacterial pathogens. Overall, it discovered agents that block TonB-dependent transport and showed the potential to find compounds that act against Gram-negative CRE (carbapenem-resistant Enterobacteriaceae)/ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) pathogens. Our results suggest that hundreds of such chemicals may exist in larger compound libraries.IMPORTANCE Antibiotic resistance in Gram-negative bacteria has spurred efforts to find novel compounds against new targets. The CRE/ESKAPE pathogens are resistant bacteria that include Acinetobacter baumannii, a common cause of ventilator-associated pneumonia and sepsis. We performed fluorescence high-throughput screening (FLHTS) against Escherichia coli to find inhibitors of TonB-dependent iron transport, tested them against A. baumannii, and then adapted the FLHTS technology to allow direct screening against A. baumannii This methodology is expandable to other drug-resistant Gram-negative pathogens. Compounds that block TonB action may interfere with iron acquisition from eukaryotic hosts and thereby constitute bacteriostatic antibiotics that prevent microbial colonization of human and animals. The FLHTS method may identify both species-specific and broad-spectrum agents against Gram-negative bacteria.
journal_name
J Bacterioljournal_title
Journal of bacteriologyauthors
Nairn BL,Eliasson OS,Hyder DR,Long NJ,Majumdar A,Chakravorty S,McDonald P,Roy A,Newton SM,Klebba PEdoi
10.1128/JB.00889-16subject
Has Abstractpub_date
2017-04-25 00:00:00issue
10eissn
0021-9193issn
1098-5530pii
JB.00889-16journal_volume
199pub_type
杂志文章abstract::The P1 plasmid addiction operon is a compact genetic structure consisting of promoter, operator, antitoxin gene (phd), and toxin gene (doc). The 73-amino-acid antitoxin protein, Phd, has two distinct functions: it represses transcription (by binding to its operator) and it prevents host death (by binding and neutraliz...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.186.9.2692-2698.2004
更新日期:2004-05-01 00:00:00
abstract::Hall, Herbert E. (Robert A. Taft Sanitary Engineering Center, Cincinnati, Ohio), Robert Angelotti, Keith H. Lewis, and Milton J. Foter. Characteristics of Clostridium perfringens strains associated with food and food-borne disease. J. Bacteriol. 85:1094-1103. 1963.-A total of 83 strains of Clostridium perfringens-30 f...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.85.5.1094-1103.1963
更新日期:1963-05-01 00:00:00
abstract::The citrate transporter CitP of lactic acid bacteria catalyzes electrogenic precursor-product exchange of citrate versus L-lactate during citrate-glucose cometabolism. In the absence of sugar, L-lactate is replaced by the metabolic intermediates/end products pyruvate, α-acetolactate, and acetate. In this study, the bi...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.00196-12
更新日期:2012-07-01 00:00:00
abstract::Most mutations in the yajC-secDF operon identified via genetic screens confer a cold-sensitive growth phenotype. Here we report that two of these mutations confer this cold-sensitive phenotype by inactivating the SecDF-YajC complex in protein translocation. ...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.187.19.6852-6855.2005
更新日期:2005-10-01 00:00:00
abstract::The PriA protein, a component of the phiX174-type primosome, was previously shown to be essential for damage-inducible DNA replication in Escherichia coli, termed inducible stable DNA replication. Here, we show that priA::kan null mutants are defective in transductional and conjugational homologous recombination and a...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.178.5.1258-1264.1996
更新日期:1996-03-01 00:00:00
abstract::A monoclonal antibody (MAb), MAb 900, which detects a 43-kDa protein present on Escherichia coli was found. Subsequently, more than 90 organisms, belonging to either the bacterial, archaeal, or eucaryal domain, were tested for reactivity to this MAb. Of the bacterial and archaeal domains, almost all species proved to ...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.177.1.11-19.1995
更新日期:1995-01-01 00:00:00
abstract::The radB101 and recN262 mutations showed essentially identical phenotypes when compared in isogenic Escherichia coli strains for their effects on gamma and UV radiation survival and on conjugal recombination in a uvrA recB recC sbcB sbcC strain. Complementation tests involving attempts to reconstitute a radB+ recN+ st...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.170.5.2392-2394.1988
更新日期:1988-05-01 00:00:00
abstract::We examined the kinetics of beta-galactosidase (EC 3.2.1.23) induction in the yeast Kluyveromyces lactis. Enzyme activity began to increase 10 to 15 min, about 1/10 of a cell generation, after the addition of inducer and continued to increase linearly for from 7 to 9 cell generations before reaching a maximum, some 12...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.142.3.777-785.1980
更新日期:1980-06-01 00:00:00
abstract::Lambda transducing phage gamma 16-2 carries the genes envA, ftsZ, ftsA, ddl, and murC and directs the synthesis of six unique proteins in ultraviolet-irradiated cells. Various derivatives of gamma 16-2 carrying smaller segments of the bacterial deoxyribonucleic acid have also been analyzed for their capacity to direct...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.143.3.1281-1288.1980
更新日期:1980-09-01 00:00:00
abstract::A mutant of Mycobacterium smegmatis unable to use the dipeptide carnosine (beta-alanyl-L-histidine) as a sole carbon or nitrogen source was isolated. Carnosinase activity and the ability to grow on beta-Ala and/or L-His were similar in the mutant and the wild type. However, the mutant showed significant impairment in ...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.180.24.6773-6775.1998
更新日期:1998-12-01 00:00:00
abstract::FabG performs the NADPH-dependent reduction of β-keto acyl-acyl carrier protein substrates in the elongation cycle of fatty acid synthesis. We report the characterization of a temperature-sensitive mutation (fabGΔ8) in Escherichia colifabG that results from an in-frame 8-amino-acid residue deletion in the α6/α7 subdom...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.00074-17
更新日期:2017-04-25 00:00:00
abstract::Target analyses of the induction of rho(-) mutants indicated that the cytoplasm of an aerobic nonrepressed yeast contains approximately 20 mutable units, whereas repressed cells exhibit approximately 3. Aerobic adaptation of fully repressed cells brings about an increase in these cytoplasmic units which continues afte...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.97.3.1142-1148.1969
更新日期:1969-03-01 00:00:00
abstract::Mycobacterium avium-Mycobacterium intracellulare complex (MAC) is the most common isolate of nontuberculous mycobacteria and causes pulmonary and extrapulmonary diseases. MAC species can be grouped into 31 serotypes by the epitopic oligosaccharide structure of the species-specific glycopeptidolipid (GPL) antigen. The ...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.01850-07
更新日期:2008-05-01 00:00:00
abstract::In Neurospora crassa, the expression of nit-3, the structural gene which encodes nitrate reductase, is highly regulated and requires both nitrate induction and nitrogen catabolite derepression. The major nitrogen regulatory gene, nit-2, acts in a positive fashion to turn on the expression of nit-3 and other nitrogen-r...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.170.2.657-661.1988
更新日期:1988-02-01 00:00:00
abstract::Functional domain structures of pyocins AP41, S1, and S2 were assigned by examining the functions of chimeric pyocins and deletion derivatives. Pyocins AP41, S1, and S2 are essentially composed of three domains, the receptor-binding domain, the translocation domain, and the DNase domain, in that order from the N termi...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.175.19.6179-6185.1993
更新日期:1993-10-01 00:00:00
abstract::Many surface proteins of pathogenic gram-positive bacteria are linked to the cell wall envelope by a mechanism requiring a C-terminal sorting signal with an LPXTG motif. Surface proteins of Streptococcus pneumoniae harbor another motif, YSIRK-G/S, which is positioned within signal peptides. The signal peptides of some...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.185.9.2910-2919.2003
更新日期:2003-05-01 00:00:00
abstract::Korormicin is an antibiotic produced by some pseudoalteromonads which selectively kills Gram-negative bacteria that express the Na+-pumping NADH:quinone oxidoreductase (Na+-NQR.) We show that although korormicin is an inhibitor of Na+-NQR, the antibiotic action is not a direct result of inhibiting enzyme activity. Ins...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.00718-18
更新日期:2019-05-08 00:00:00
abstract::The interaction of Streptococcus pyogenes (group A streptococcus [GAS]) with its human host requires several surface proteins. In this study, we isolated mutations in a gene required for the surface localization of protein F by transposon mutagenesis of the M6 strain JRS4. This gene (srtA) encodes a protein homologous...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.184.8.2181-2191.2002
更新日期:2002-04-01 00:00:00
abstract::A sucrose-inducible alpha-glucosidase activity that hydrolyzes sucrose in Candida albicans has been demonstrated previously. The enzyme is assayable in whole cells and was inhibited by both sucrose and maltose. A C. albicans gene (CASUC1) that affects sucrose utilization and alpha-glucosidase activity was cloned by ex...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.174.1.222-232.1992
更新日期:1992-01-01 00:00:00
abstract::DNA was prepared from a strain of Escherichia coli bearing a mutation which confers the GlnC phenotype (inability to reduce the expression of glnA and other nitrogen-regulated operons in response to ammonia in the growth medium). A fragment of this DNA carrying glnA, the structural gene for glutamine synthetase, was c...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.150.1.214-220.1982
更新日期:1982-04-01 00:00:00
abstract::We have identified three new Haemophilus influenzae mutations causing cells to exhibit extreme hypercompetence at all stages of growth. The mutations are in murE, which encodes the meso-diaminopimelate-adding enzyme of peptidoglycan synthesis. All are point mutations causing nonconservative amino acid substitutions, t...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.182.12.3323-3330.2000
更新日期:2000-06-01 00:00:00
abstract::The fec region of the Escherichia coli chromosome determines a citrate-dependent iron(III) transport system. The nucleotide sequence of fec revealed five genes, fecABCDE, which are transcribed from fecA to fecE. The fecA gene encodes a previously described outer membrane receptor protein. The fecB gene product is form...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.171.5.2626-2633.1989
更新日期:1989-05-01 00:00:00
abstract::Brown, Eric R. (Northwestern University Medical School, Chicago, Ill.), Peter Buinauskas, and Steven O. Schwartz. Immunofluorescent antibody studies of a murine leukemia virus. J. Bacteriol. 92:978-982. 1966.-Correlation was close between in vitro complement fixation, immunodiffusion, and passive cutaneous anaphylaxis...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.92.4.978-982.1966
更新日期:1966-10-01 00:00:00
abstract::The peptidoglycan from two types of filamentous cell division mutants of Agmenellum quadruplicatum strain BG1 has been compared to that of the parent organism. Small variations in the total peptidoglycan composition on a dry-weight basis were found in the mutants. The reduced level of peptidoglycan in the serpentine m...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.111.2.614-621.1972
更新日期:1972-08-01 00:00:00
abstract::Previously, the effects of three point mutations (at amino acid positions 31, 84, and 194) in the gene coding for a sulfonamide-resistant dihydropteroate synthase of Neisseria meningitidis were analyzed by site-directed mutagenesis. Changes at positions 31 and 194 abolished the phenotypic expression of sulfonamide res...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.179.3.831-837.1997
更新日期:1997-02-01 00:00:00
abstract::The effects of surfaces on the physiology of bacteria adhering to surfaces or immobilized within biofilms are receiving more interest. A study of the effects of hydrophobic and hydrophilic substrata on the colonization behavior of a marine bacterium, SW5, revealed major differences in the morphology of SW5 on these su...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.176.22.6900-6906.1994
更新日期:1994-11-01 00:00:00
abstract::We have overexpressed and purified the Helicobacter pylori Fur protein and analyzed its interaction with the intergenic regions of divergent genes involved in iron uptake (frpB and ceuE) and oxygen radical detoxification (katA and tsaA). DNase I footprint analysis showed that Fur binds specifically to a high-affinity ...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.183.16.4932-4937.2001
更新日期:2001-08-01 00:00:00
abstract::The temperature-sensitive cyr1-2 mutant in Saccharomyces cerevisiae produces low levels of adenylate cyclase and cyclic AMP at 25 degrees C and is unable to synthesize repressible acid phosphatase at 25 degrees C. Suppressor mutants of cyr1-2 were isolated by detecting acid phosphatase activity. One of the dominant su...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/jb.173.7.2406-2408.1991
更新日期:1991-04-01 00:00:00
abstract::The survival of ultraviolet (UV)-irradiated cultures of Haemophilus influenzae Rd is determined by at least two responses: (i) excision-repair ability and (ii) UV-induced cell lysis. An UV-resistant mutant, BC200, has the same capabilities as the wild type, Rd, for excising dimers but does not exhibit lysis. Lytic res...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.103.1.9-15.1970
更新日期:1970-07-01 00:00:00
abstract::A lipopolysaccharide (LPS) has been isolated from the gram-negative photosynthetic bacterium Rhodopseudomonas capsulata. Chemical analysis revealed the presence of d-glucose, d-galactose, l-rhamnose, 3-O-methyl-l-rhamnose (l-acofriose), d-glucosamine, 2-keto-3-deoxyoctonate, and neuraminic acid. The LPS does not conta...
journal_title:Journal of bacteriology
pub_type: 杂志文章
doi:10.1128/JB.109.3.1106-1113.1972
更新日期:1972-03-01 00:00:00