Dataset Information

Proteomic analysis of the papaverine degrading Arthrobacter sp.

ABSTRACT: In previous work in our group, shotgun genome sequencing of Arthrobacter sp. revealed potential new P450 monooxygenases and many other oxidoreductases with putative hydroxylation activity. A targeted approach to identify enzymes involved in the degradation of certain molecules is proteomic analysis. In the case of growth on certain substances, enzymes like P450s, which are responsible for the observed organism’s capabilities, might be overexpressed or initially induced.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Arthrobacter Sp. Hg

SUBMITTER: Jan Klenk

LAB HEAD: Bernhard Hauer

PROVIDER: PXD009566 | Pride | 2019-02-19

REPOSITORIES: Pride

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Publications

Identification and characterization of cytochrome P450 1232A24 and 1232F1 from Arthrobacter sp. and their role in the metabolic pathway of papaverine.

Klenk Jan M JM Fischer Max-Philipp MP Dubiel Paulina P Sharma Mahima M Rowlinson Benjamin B Grogan Gideon G Hauer Bernhard B

Journal of biochemistry 20190701 1

Cytochrome P450 monooxygenases (P450s) play crucial roles in the cell metabolism and provide an unsurpassed diversity of catalysed reactions. Here, we report the identification and biochemical characterization of two P450s from Arthrobacter sp., a Gram-positive organism known to degrade the opium alkaloid papaverine. Combining phylogenetic and genomic analysis suggested physiological roles for P450s in metabolism and revealed potential gene clusters with redox partners facilitating the reconstit ...[more]

PMID: 30759214

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Project description:Pneumonia is a serious problem worldwide. We recently demonstrated that innate defense mechanisms of the lung are highly inducible against pneumococcal pneumonia. To determine the breadth of protection conferred by stimulation of lung mucosal innate immunity, and to identify cells and signaling pathways activated by this treatment, mice were treated with an aerosolized bacterial lysate, then challenged with lethal doses of bacterial and fungal pathogens. Mice were highly protected against a broad array of Gram-positive, Gram-negative, and Class A bioterror bacterial pathogens, and Aspergillus fumigatus. Protection was associated with rapid pathogen killing within the lungs, and this effect was recapitulated in vitro using a respiratory epithelial cell line. Gene expression analysis of lung tissue showed marked activation of NF-kappaB, Type I and II interferon, and antifungal Card9-Bcl10-Malt1 pathways. Cytokines were the most strongly induced genes, but the inflammatory cytokines TNF and IL-6 were not required for protection. Lung-expressed antimicrobial peptides were also highly upregulated. Taken together, stimulated innate resistance (StIR) appears to occur through the activation of multiple host defense signaling pathways in lung epithelial cells, inducing rapid pathogen killing, and conferring broad protection against virulent bacterial and fungal pathogens. Augmentation of innate antimicrobial defenses of the lungs might have therapeutic value for protection of patients with neutropenia or impaired adaptive immunity against opportunistic pneumonia, and for defense of immunocompetent subjects against a bioterror threat or epidemic respiratory infection. Keywords: differential gene expression; time course; innate immunity; pneumonia; immunocompromised host; lung epithelium Gene expression patterns in mouse lung homogenates were analyzed 2h after exposure to aerosolized PBS (Sham treatment), 2h after exposure to aerosolized NTHi lysate or 4h after exposure to aerosolized NTHi lysate. Each group consisted of six mice.

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Dataset Information

Proteomic analysis of the papaverine degrading Arthrobacter sp.

Publications

Identification and characterization of cytochrome P450 1232A24 and 1232F1 from Arthrobacter sp. and their role in the metabolic pathway of papaverine.

Similar Datasets

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