Article

Disulfiram irreversibly aggregates betaine aldehyde dehydrogenase--a potential target for antimicrobial agents against Pseudomonas aeruginosa.

Laboratorio de Osmorregulación, Facultad de Estudios Superiores Iztacala, Universidad Nacional Autónoma de México, Avenida de los Barrios, Tlalnepantla, Estado de México, 54090 México, Mexico.
Biochemical and Biophysical Research Communications (Impact Factor: 2.28). 04/2006; 341(2):408-15. DOI: 10.1016/j.bbrc.2006.01.003
Source: PubMed

ABSTRACT In the human pathogen Pseudomonas aeruginosa, betaine aldehyde dehydrogenase (PaBADH) may play the dual role of assimilating carbon and nitrogen from choline or choline precursors--abundant at infection sites--and producing glycine betaine, which protects the bacterium against the high-osmolality stress prevalent in the infected tissues. This tetrameric enzyme contains four cysteine residues per subunit and is a potential drug target. In our search for specific inhibitors, we mutated the catalytic Cys286 to alanine and chemically modified the recombinant wild-type and the four Cys-->Ala single mutants with thiol reagents. The small methyl-methanethiosulfonate inactivated the enzymes without affecting their stability while the bulkier dithionitrobenzoic acid (DTNB) and bis[diethylthiocarbamyl] disulfide (disulfiram) induced enzyme dissociation--at 23 degrees C--and irreversible aggregation--at 37 degrees C. Of the four Cys-->Ala mutants only C286A retained its tetrameric structure after DTNB or disulfiram treatments, suggesting that steric constraints arising upon the covalent attachment of a bulky group to C286 resulted in distortion of the backbone configuration in the active site region followed by a severe decrease in enzyme stability. Since neither NAD(P)H nor betaine aldehyde prevented disulfiram-induced PaBADH inactivation or aggregation, and reduced glutathione was unable to restore the activity of the modified enzyme, we propose that disulfiram could be a useful drug to combat infection by P. aeruginosa.

0 Bookmarks
 · 
81 Views
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Members of the Burkholderia cepacia complex (Bcc) are important in medical, biotechnological, and agricultural disciplines. These bacteria naturally occur in soil and water environments and have adapted to survive in association with plants and animals including humans. All Bcc species are opportunistic pathogens including Burkholderia cenocepacia that causes infections in cystic fibrosis and chronic granulomatous disease patients. The adaptation of B. cenocepacia to the host environment was assessed in a rat chronic respiratory infection model and compared to that of high cell-density in vitro grown cultures using transcriptomics. The distribution of genes differentially expressed on chromosomes 1, 2, and 3 was relatively proportional to the size of each genomic element, whereas the proportion of plasmid-encoded genes differentially expressed was much higher relative to its size and most genes were induced in vivo. The majority of genes encoding known virulence factors, components of types II and III secretion systems and chromosome 2-encoded type IV secretion system were similarly expressed between in vitro and in vivo environments. Lower expression in vivo was detected for genes encoding N-acyl-homoserine lactone synthase CepI, orphan LuxR homolog CepR2, zinc metalloproteases ZmpA and ZmpB, LysR-type transcriptional regulator ShvR, nematocidal protein AidA, and genes associated with flagellar motility, Flp type pilus formation, and type VI secretion. Plasmid-encoded type IV secretion genes were markedly induced in vivo. Additional genes induced in vivo included genes predicted to be involved in osmotic stress adaptation or intracellular survival, metal ion, and nutrient transport, as well as those encoding outer membrane proteins. Genes identified in this study are potentially important for virulence during host-pathogen interactions and may be associated with survival and adaptation to the host environment during chronic lung infections.
    Frontiers in Cellular and Infection Microbiology 01/2011; 1:15. · 2.62 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: The purpose of the present research was to evaluate the efficacy of problem solving workshops designed for parents of pre-teens. In these workshops topics such as understanding others’ emotions, ideas, and motivations, as well as finding various solutions for a problem, were taught. Participants included 96 mothers of students in private schools. These mothers were instructed in five workshops lasted ten sessions, two hours each. Three subscales of Family Assessment Device (FAD) were used before and after the workshops: communication, affective responsiveness, and affective involvement. Analysis of data indicates that the workshops had positive effects on subjects’ performance in “affective involvement”. © 2011 Published by Elsevier Ltd.
    Procedia - Social and Behavioral Sciences 01/2011; 15:957-960.
  • [Show abstract] [Hide abstract]
    ABSTRACT: Mangroves are salt-tolerant forest ecosystem that extends between tropical and subtropical intertidal regions of the world. Mangroves are biochemically unique vegetation that produce wide array of natural products with immense medicinal potential. These plants are the most valuable resources and provide economic and ecological benefits to the coastal people. Natural products from these plants are of great interest as they provide innumerable direct and indirect benefits to human beings for the discovery of novel antimicrobial and other bioactive compounds. They possess active metabolites with some novel chemical structures that belong to diverse chemical classes such as alkaloids, phenol, steroids, terpenoids and tannins. Several mangrove species have been used in traditional medicine or have few applications as insecticide and pesticide. To date, several mangroves, and their associated species and solvent extracts are screened for antimicrobial activity along with the presence of potent bioactive compounds. The present article emphasizes and creates awareness about the potential mangrove plants and their associates as a source of biologically active compounds with potent antimicrobial properties. This paper also elaborates the mechanisms of action and various methods for screening of antimicrobial compounds.
    Chinese Journal of Integrative Medicine 01/2014; · 1.06 Impact Factor