In Vitro Biochemical Study of CYP51-Mediated Azole Resistance in Aspergillus fumigatus.

Abstract:

:The incidence of triazole-resistant Aspergillus infections is increasing worldwide, often mediated through mutations in the CYP51A amino acid sequence. New classes of azole-based drugs are required to combat the increasing resistance to existing triazole therapeutics. In this study, a CYP51 reconstitution assay is described consisting of eburicol, purified recombinant Aspergillus fumigatus CPR1 (AfCPR1), and Escherichia coli membrane suspensions containing recombinant A. fumigatus CYP51 proteins, allowing in vitro screening of azole antifungals. Azole-CYP51 studies determining the 50% inhibitory concentration (IC50) showed that A. fumigatus CYP51B (Af51B IC50, 0.50 μM) was 34-fold more susceptible to inhibition by fluconazole than A. fumigatus CYP51A (Af51A IC50, 17 μM) and that Af51A and Af51B were equally susceptible to inhibition by voriconazole, itraconazole, and posaconazole (IC50s of 0.16 to 0.38 μM). Af51A-G54W and Af51A-M220K enzymes were 11- and 15-fold less susceptible to inhibition by itraconazole and 30- and 8-fold less susceptible to inhibition by posaconazole than wild-type Af51A, confirming the azole-resistant phenotype of these two Af51A mutations. Susceptibility to voriconazole of Af51A-G54W and Af51A-M220K was only marginally lower than that of wild-type Af51A. Susceptibility of Af51A-L98H to inhibition by voriconazole, itraconazole, and posaconazole was only marginally lower (less than 2-fold) than that of wild-type Af51A. However, Af51A-L98H retained 5 to 8% residual activity in the presence of 32 μM triazole, which could confer azole resistance in A. fumigatus strains that harbor the Af51A-L98H mutation. The AfCPR1/Af51 assay system demonstrated the biochemical basis for the increased azole resistance of A. fumigatus strains harboring G54W, L98H, and M220K Af51A point mutations.

authors

Warrilow AG,Parker JE,Price CL,Nes WD,Kelly SL,Kelly DE

doi

10.1128/AAC.01806-15

subject

Has Abstract

pub_date

2015-12-01 00:00:00

pages

7771-8

issue

12

eissn

0066-4804

issn

1098-6596

pii

AAC.01806-15

journal_volume

59

pub_type

杂志文章
  • Interplay between Antibiotic Efficacy and Drug-Induced Lysis Underlies Enhanced Biofilm Formation at Subinhibitory Drug Concentrations.

    abstract::Subinhibitory concentrations of antibiotics have been shown to enhance biofilm formation in multiple bacterial species. While antibiotic exposure has been associated with modulated expression of many biofilm-related genes, the mechanisms of drug-induced biofilm formation remain a focus of ongoing research efforts and ...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/AAC.01603-17

    authors: Yu W,Hallinen KM,Wood KB

    更新日期:2017-12-21 00:00:00

  • Azasordarins: susceptibility of fluconazole-susceptible and fluconazole-resistant clinical isolates of Candida spp. to GW 471558.

    abstract::The in vitro activity of the azasordarin GW 471558 was compared with those of amphotericin B, flucytosine, itraconazole, and ketoconazole against 177 clinical isolates of Candida spp. GW 471558 showed potent activity against Candida albicans, Candida glabrata, and Candida tropicalis, even against isolates with decreas...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/AAC.45.6.1905-1907.2001

    authors: Cuenca-Estrella M,Mellado E,Díaz-Guerra TM,Monzón A,Rodríguez-Tudela JL

    更新日期:2001-06-01 00:00:00

  • Efficacy of tigecycline for secondary Acinetobacter bacteremia and factors associated with treatment failure.

    abstract::We describe the clinical outcome of 17 patients with secondary Acinetobacter bacteremia whose isolates had a tigecycline MIC of ≤2 mg/liter and who received tigecycline within 2 days of bacteremia onset. The 14-day mortality rate of the tigecycline cohort was 41.2% (7/17), which was significantly higher than that of t...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/AAC.04987-14

    authors: Liou BH,Lee YT,Kuo SC,Liu PY,Fung CP

    更新日期:2015-01-01 00:00:00

  • NorM, a putative multidrug efflux protein, of Vibrio parahaemolyticus and its homolog in Escherichia coli.

    abstract::We found that cells of Vibrio parahaemolyticus possess an energy-dependent efflux system for norfloxacin. We cloned a gene for a putative norfloxacin efflux protein from the chromosomal DNA of V. parahaemolyticus by using an Escherichia coli mutant lacking the major multidrug efflux system AcrAB as the host and sequen...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/AAC.42.7.1778

    authors: Morita Y,Kodama K,Shiota S,Mine T,Kataoka A,Mizushima T,Tsuchiya T

    更新日期:1998-07-01 00:00:00

  • Evolution of a 72-Kilobase Cointegrant, Conjugative Multiresistance Plasmid in Community-Associated Methicillin-Resistant Staphylococcus aureus Isolates from the Early 1990s.

    abstract::Horizontal transfer of plasmids encoding antimicrobial resistance and virulence determinants has been instrumental in Staphylococcus aureus evolution, including the emergence of community-associated methicillin-resistant S. aureus (CA-MRSA). In the early 1990s, the first CA-MRSA strain isolated in Western Australia (W...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/AAC.01560-19

    authors: Yui Eto K,Firth N,Davis AM,Kwong SM,Krysiak M,Lee YT,O'Brien FG,Grubb WB,Coombs GW,Bond CS,Ramsay JP

    更新日期:2019-10-22 00:00:00

  • Evaluation of the Efficacy of ME1111 in the Topical Treatment of Dermatophytosis in a Guinea Pig Model.

    abstract::The treatment of dermatophytoses, including onychomycosis, has come a long way over the past few decades with the introduction of oral antifungals (e.g., terbinafine and itraconazole). However, with these advancements in oral therapies come several undesirable effects, such as kidney and liver toxicity, along with dru...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/AAC.03073-15

    authors: Long L,Hager C,Ghannoum M

    更新日期:2016-03-25 00:00:00

  • Genomic analysis of the multidrug-resistant Acinetobacter baumannii strain MDR-ZJ06 widely spread in China.

    abstract::We previously reported that the multidrug-resistant (MDR) Acinetobacter baumannii strain MDR-ZJ06, belonging to European clone II, was widely spread in China. In this study, we report the whole-genome sequence of this clinically important strain. A 38.6-kb AbaR-type genomic resistance island (AbaR22) was identified in...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/AAC.01134-10

    authors: Zhou H,Zhang T,Yu D,Pi B,Yang Q,Zhou J,Hu S,Yu Y

    更新日期:2011-10-01 00:00:00

  • Paradoxical effect of clindamycin in experimental, acute toxoplasmosis in cats.

    abstract::Cats were experimentally inoculated parenterally with the ME49 strain of Toxoplasma gondii to characterize the efficacies of two different dosages of orally administered clindamycin hydrochloride in the treatment of ocular toxoplasmosis. Concentrations of clindamycin hydrochloride at levels previously suggested to be ...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/AAC.40.6.1352

    authors: Davidson MG,Lappin MR,Rottman JR,Tompkins MB,English RV,Bruce AT,Jayawickrama J

    更新日期:1996-06-01 00:00:00

  • beta-lactamase production and antimicrobial susceptibility of oral heterogeneous Fusobacterium nucleatum populations in young children.

    abstract::Oral Fusobacterium nucleatum populations from 20 young, healthy children were examined for beta-lactamase production. Ten children (50%) harbored, altogether, 25 beta-lactamase-positive F. nucleatum isolates that were identified as F. nucleatum subsp. polymorphum, F. nucleatum subsp. nucleatum, and F. nucleatum subsp....

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/AAC.43.5.1270

    authors: Könönen E,Kanervo A,Salminen K,Jousimies-Somer H

    更新日期:1999-05-01 00:00:00

  • Structure-absorption relationships of a series of 6-fluoroquinolones.

    abstract::The physicochemical constants and some structural parameters (topological, steric, and electronic) of eight third-generation monofluorate quinolones (six uncommercialized and two used clinically [ciprofloxacin and enrofloxacin]) were determined: pKa, intrinsic solubility (S0), chromatographic capacity factor, partitio...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/AAC.41.9.1996

    authors: Escribano E,Calpena AC,Garrigues TM,Freixas J,Domenech J,Moreno J

    更新日期:1997-09-01 00:00:00

  • Evolution and global dissemination of macrolide-resistant group A streptococci.

    abstract::Macrolide-resistant group A streptococci (MRGAS) have been recovered from many countries worldwide. However, the strain typing information that is available has been insufficient for estimating the total number of macrolide-resistant clones, their geographic distributions, and their evolutionary relationships. In this...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/AAC.00325-06

    authors: Robinson DA,Sutcliffe JA,Tewodros W,Manoharan A,Bessen DE

    更新日期:2006-09-01 00:00:00

  • Synergism between penicillin, clindamycin, or metronidazole and gentamicin against species of the Bacteroides melaninogenicus and Bacteroides fragilis groups.

    abstract::Clinical isolates of the Bacteroides melaninogenicus and Bacteroides fragilis groups were tested for in vitro and in vivo susceptibility to penicillin, clindamycin, and metronidazole, used singly or in combination with gentamicin. The in vitro tests consisted of determinations of minimal inhibitory concentrations (MIC...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/aac.25.1.71

    authors: Brook I,Coolbaugh JC,Walker RI,Weiss E

    更新日期:1984-01-01 00:00:00

  • Interaction of metronidazole with resistant and susceptible Bacteroides fragilis.

    abstract::The kinetics of the lethal action of metronidazole and the formation of acetamide have been studied in a strain of Bacteroides fragilis which is relatively resistant to metronidazole. As with a susceptible strain of B. fragilis, the data are consistent with a model in which a labile intermediate in metronidazole metab...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/aac.21.1.131

    authors: McLafferty MA,Koch RL,Goldman P

    更新日期:1982-01-01 00:00:00

  • Double-blind clinical trials of oral cyclacillin and ampicillin.

    abstract::A double-blind study was performed to compare the clinical response and the incidence of side effects in 2,581 patients administered ampicillin or cyclacillin for infections of the genitourinary or respiratory tract, infections of the skin and soft tissues, or for otitis media. There was no significant difference in c...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 临床试验,杂志文章,随机对照试验

    doi:10.1128/aac.15.1.55

    authors: Gold JA,Hegarty CP,Deitch MW,Walker BR

    更新日期:1979-01-01 00:00:00

  • In vitro activity of RP 74501-RP 74502, a novel streptogramin antimicrobial mixture, against clinical isolates of Legionella species.

    abstract::Agar and broth microdilution MICs of RP 74501-RP 74502, a mixture of streptogramin antimicrobial agents that inhibited 90% of 22 Legionella strains tested, were 0.64 and 0.08 microgram/ml, respectively; respective erythromycin values were 1.0 and 0.12 microgram/ml. RP 74501-RP 74502 at 1 microgram/ml was more active t...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/aac.37.4.908

    authors: Edelstein PH,Edelstein MA

    更新日期:1993-04-01 00:00:00

  • Antibiotic resistance and substrate profiles of the class A carbapenemase KPC-6.

    abstract::The class A carbapenemase KPC-6 produces resistance to a broad range of β-lactam antibiotics. This enzyme hydrolyzes penicillins, the monobactam aztreonam, and carbapenems with similar catalytic efficiencies, ranging from 10(5) to 10(6) M(-1) s(-1). The catalytic efficiencies of KPC-6 against cephems vary to a greater...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/AAC.01338-12

    authors: Lamoureaux TL,Frase H,Antunes NT,Vakulenko SB

    更新日期:2012-11-01 00:00:00

  • Growth of Physarum gyrosum on agar plates and in liquid culture.

    abstract::The physical and nutritional requirements of the antibiotic-producing slime mold Physarum gyrosum were examined to develop a liquid medium for this myxomycete. Liquid culture is desired to expedite a useful scale of production of antibiotic materials for ease of isolation and structure study. Culture conditions were s...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/aac.10.4.613

    authors: Taylor RL,Mallette MF

    更新日期:1976-10-01 00:00:00

  • Antimicrobial susceptibility among clinical Nocardia species identified by multilocus sequence analysis.

    abstract::Antimicrobial susceptibility patterns of 112 clinical isolates, 28 type strains, and 9 reference strains of Nocardia were determined using the Sensititre Rapmyco microdilution panel (Thermo Fisher, Inc.). Isolates were identified by highly discriminatory multilocus sequence analysis and were chosen to represent the di...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/AAC.02770-14

    authors: McTaggart LR,Doucet J,Witkowska M,Richardson SE

    更新日期:2015-01-01 00:00:00

  • Synthesis and antibacterial activities of optically active ofloxacin.

    abstract::Two optically active (100% enantiomeric excess) isomers of ofloxacin [(+/-)-ofloxacin; DL-8280; (+/-)-9-fluoro-2,3-dihydro-3-methyl-10-(4-methyl-1-piperazinyl)-7-oxo-7 H-pyrido[1,2,3-de] [1,4] benzoxazine-6-carboxylic acid] were prepared by use of their optically resolved synthetic intermediates. One of the isomers, (...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/aac.29.1.163

    authors: Hayakawa I,Atarashi S,Yokohama S,Imamura M,Sakano K,Furukawa M

    更新日期:1986-01-01 00:00:00

  • Development of rifapentine susceptibility tests for Mycobacterium tuberculosis.

    abstract::Two methods for testing the susceptibility of Mycobacterium tuberculosis to rifapentine have been developed: the agar proportion method and the radiometric BACTEC technique. A critical concentration of 0.5 microg of rifapentine per ml is proposed for both methods since it provides a reliable means of distinguishing be...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/AAC.43.1.25

    authors: Heifets L,Sanchez T,Vanderkolk J,Pham V

    更新日期:1999-01-01 00:00:00

  • Absence of synergistic activity between ampicillin and vancomycin against highly vancomycin-resistant enterococci.

    abstract::The emergence of clinical enterococcal isolates resistant to both ampicillin and vancomycin is a cause of great concern, as there are few therapeutic alternatives for treatment of infections caused by such organisms. We evaluated the effects of the combination of ampicillin with vancomycin against vancomycin-resistant...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/aac.36.10.2201

    authors: Cercenado E,Eliopoulos GM,Wennersten CB,Moellering RC Jr

    更新日期:1992-10-01 00:00:00

  • Penetration of cefprozil into middle ear fluid of patients with otitis media.

    abstract::Penetration of cefprozil into the middle ear fluid was investigated in patients with chronic otitis media. A total of 89 patients ranging from 7 months to 11 years old participated in the study. The middle ear fluid was removed by ventilation tubes inserted through the tympanic membrane at times ranging from 0.38 to 5...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 临床试验,杂志文章,随机对照试验

    doi:10.1128/aac.38.9.2210

    authors: Shyu WC,Haddad J,Reilly J,Khan WN,Campbell DA,Tsai Y,Barbhaiya RH

    更新日期:1994-09-01 00:00:00

  • Efficacy of monoclonal antibodies against defined antigens for passive immunotherapy of chronic gastrointestinal cryptosporidiosis.

    abstract::Cryptosporidium parvum is an important cause of diarrhea in humans and calves and can persistently infect immunocompromised hosts. Presently, there are no consistently effective parasite-specific drugs for cryptosporidiosis. We hypothesized that neutralizing monoclonal antibodies (MAbs) targeting the apical complex an...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/aac.46.2.275-282.2002

    authors: Riggs MW,Schaefer DA,Kapil SJ,Barley-Maloney L,Perryman LE

    更新日期:2002-02-01 00:00:00

  • Killing and lysis of Echerichia coli in the presence of choloramphenicol: relation to cellular magensim.

    abstract::Treatment of Escherichia coli K-10 with 100 mug of chloramphenicol per ml for periods greater than 30 min leads to progressive lysis and killing of cells. The bactericidal action of the antibiotic is dependent on cell growth and physiology; only rapidly dividing cells are susceptible to killing; resting or slowly grow...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/aac.7.6.748

    authors: Guota RS

    更新日期:1975-06-01 00:00:00

  • Implications of Antimicrobial Combinations in Complex Wound Biofilms Containing Fungi.

    abstract::Diabetic foot ulcer treatment currently focuses on targeting bacterial biofilms, while dismissing fungi. To investigate this, we used an in vitro biofilm model containing bacteria and fungi, reflective of the wound environment, to test the impact of antimicrobials. Here we showed that while monotreatment approaches in...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/AAC.00672-17

    authors: Townsend EM,Sherry L,Kean R,Hansom D,Mackay WG,Williams C,Butcher J,Ramage G

    更新日期:2017-08-24 00:00:00

  • Analysis of amifloxacin in plasma and urine by high-pressure liquid chromatography and intravenous pharmacokinetics in rhesus monkeys.

    abstract::An analytical method for the quantitation of amifloxacin, 6-fluoro-1,4-dihydro-1-(methylamino)-7-(4-methyl-1-piperazinyl)-4-oxo-3- quinolinecarboxylic acid, in plasma and urine has been developed. The method involves extraction with chloroform, back-extraction into 0.1 M sodium hydroxide, and subsequent analysis by re...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/aac.27.5.769

    authors: McCoy LF,Crawmer BP,Benziger DP

    更新日期:1985-05-01 00:00:00

  • RND-type efflux pumps in multidrug-resistant clinical isolates of Acinetobacter baumannii: major role for AdeABC overexpression and AdeRS mutations.

    abstract::Increased expression of chromosomal genes for resistance-nodulation-cell division (RND)-type efflux systems plays a major role in the multidrug resistance (MDR) of Acinetobacter baumannii. However, the relative contributions of the three most prevalent pumps, AdeABC, AdeFGH, and AdeIJK, have not been evaluated in clin...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/AAC.02556-12

    authors: Yoon EJ,Courvalin P,Grillot-Courvalin C

    更新日期:2013-07-01 00:00:00

  • Antimicrobial therapy of experimental endocarditis caused by nutritionally variant viridans group streptococci.

    abstract::Rabbits with nutritionally variant viridans group streptococcal experimental endocarditis were treated three times daily for 3 days with procaine penicillin (1.2 X 10(6) U) alone or together with low-dose streptomycin (2 mg/kg), high-dose streptomycin (8 mg/kg), low-dose gentamicin (0.32 mg/kg), or high-dose gentamici...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/aac.30.3.465

    authors: Henry NK,Wilson WR,Roberts RB,Acar JF,Geraci JE

    更新日期:1986-09-01 00:00:00

  • Novel quinolone resistance mutations of the Escherichia coli DNA gyrase A protein: enzymatic analysis of the mutant proteins.

    abstract::Using the techniques of gap misrepair mutagenesis and site-directed mutagenesis, we have generated two novel quinolone resistance mutations of the Escherichia coli DNA gyrase A protein. DNA sequencing showed these mutations to be Ser-83----Ala and Gln-106----Arg. The mutant proteins were overproduced and purified, and...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/aac.35.2.335

    authors: Hallett P,Maxwell A

    更新日期:1991-02-01 00:00:00

  • Nucleotide sequence of the gyrA gene and characterization of ciprofloxacin-resistant mutants of Helicobacter pylori.

    abstract::PCR was used to amplify a 238-bp region from Helicobacter pylori which corresponded to the quinolone resistance-determining region in Escherichia coli. The gyrA gene of H. pylori was cloned and sequenced. An open reading frame of 2,478 nucleotides coded for a polypeptide of 826 amino acids with a calculated molecular ...

    journal_title:Antimicrobial agents and chemotherapy

    pub_type: 杂志文章

    doi:10.1128/aac.39.1.107

    authors: Moore RA,Beckthold B,Wong S,Kureishi A,Bryan LE

    更新日期:1995-01-01 00:00:00