TY - JOUR
T1 - An essential bifunctional enzyme in Mycobacterium tuberculosis for itaconate dissimilation and leucine catabolism
AU - Wang, Hua
AU - Fedorov, Alexander A.
AU - Fedorov, Elena V.
AU - Hunt, Debbie M.
AU - Rodgers, Angela
AU - Douglas, Holly L.
AU - Garza-Garcia, Acely
AU - Bonanno, Jeffrey B.
AU - Almo, Steven C.
AU - de Carvalho, Luiz Pedro Sório
N1 - Funding Information:
ACKNOWLEDGMENTS. We thank Dr. Geoff Kelly (Medical Research Council [MRC] Biomedical NMR Centre) for assistance with NMR spectroscopy. NMR data were recorded in the MRC Biomedical NMR Centre at the Francis Crick Institute, which receives core funding from Cancer Research UK Grant FC001029; Medical Research Council Grant FC001029; and Wellcome Trust Grant FC001029. Work in L.P.S.d.C.’s laboratory was supported by the Francis Crick Institute, which receives its core funding from Cancer Research UK Grant FC001060, UK MRC Grant FC001060, and Wellcome Trust Grant FC001060. L.P.S.d.C.’s laboratory also acknowledges funds from Wellcome Trust New Investigator Award 104785/B/14/Z. This research used resources of the Advanced Photon Source, a US Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract DE-AC02-06CH11357. Use of the Lilly Research Laboratories Collaborative Access Team beamline at Sector 31 of the Advanced Photon Source was provided by Eli Lilly Company, which operates the facility. The Einstein Crystallographic Core X-Ray diffraction facility is supported by NIH Shared Instrumentation Grant S10 OD020068, which we gratefully acknowledge. S.C.A. acknowledges support from the US NIH Grant P01 GM118303.
Funding Information:
We thank Dr. Geoff Kelly (Medical Research Council [MRC] Biomedical NMR Centre) for assistance with NMR spectroscopy. NMR data were recorded in the MRC Biomedical NMR Centre at the Francis Crick Institute, which receives core funding from Cancer Research UK Grant FC001029; Medical Research Council Grant FC001029; and Wellcome Trust Grant FC001029. Work in L.P.S.d.C.?s laboratory was supported by the Francis Crick Institute, which receives its core funding from Cancer Research UK Grant FC001060, UK MRC Grant FC001060, and Wellcome Trust Grant FC001060. L.P.S.d.C.?s laboratory also acknowledges funds from Wellcome Trust New Investigator Award 104785/B/14/Z. This research used resources of the Advanced Photon Source, a US Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract DE-AC02-06CH11357. Use of the Lilly Research Laboratories Collaborative Access Team beamline at Sector 31 of the Advanced Photon Source was provided by Eli Lilly Company, which operates the facility. The Einstein Crystallographic Core X-Ray diffraction facility is supported by NIH Shared Instrumentation Grant S10 OD020068, which we gratefully acknowledge. S.C.A. acknowledges support from the US NIH Grant P01 GM118303.
PY - 2019/8/6
Y1 - 2019/8/6
N2 - Mycobacterium tuberculosis (Mtb) is the etiological agent of tuberculosis. One-fourth of the global population is estimated to be infected with Mtb, accounting for ∼1.3 million deaths in 2017. As part of the immune response to Mtb infection, macrophages produce metabolites with the purpose of inhibiting or killing the bacterial cell. Itaconate is an abundant host metabolite thought to be both an antimicrobial agent and a modulator of the host inflammatory response. However, the exact mode of action of itaconate remains unclear. Here, we show that Mtb has an itaconate dissimilation pathway and that the last enzyme in this pathway, Rv2498c, also participates in L-leucine catabolism. Our results from phylogenetic analysis, in vitro enzymatic assays, X-ray crystallography, and in vivo Mtb experiments, identified Mtb Rv2498c as a bifunctional β-hydroxyacyl-CoA lyase and that deletion of the rv2498c gene from the Mtb genome resulted in attenuation in a mouse infection model. Altogether, this report describes an itaconate resistance mechanism in Mtb and an L-leucine catabolic pathway that proceeds via an unprecedented (R)-3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) stereospecific route in nature.
AB - Mycobacterium tuberculosis (Mtb) is the etiological agent of tuberculosis. One-fourth of the global population is estimated to be infected with Mtb, accounting for ∼1.3 million deaths in 2017. As part of the immune response to Mtb infection, macrophages produce metabolites with the purpose of inhibiting or killing the bacterial cell. Itaconate is an abundant host metabolite thought to be both an antimicrobial agent and a modulator of the host inflammatory response. However, the exact mode of action of itaconate remains unclear. Here, we show that Mtb has an itaconate dissimilation pathway and that the last enzyme in this pathway, Rv2498c, also participates in L-leucine catabolism. Our results from phylogenetic analysis, in vitro enzymatic assays, X-ray crystallography, and in vivo Mtb experiments, identified Mtb Rv2498c as a bifunctional β-hydroxyacyl-CoA lyase and that deletion of the rv2498c gene from the Mtb genome resulted in attenuation in a mouse infection model. Altogether, this report describes an itaconate resistance mechanism in Mtb and an L-leucine catabolic pathway that proceeds via an unprecedented (R)-3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) stereospecific route in nature.
KW - Carbon–carbon bond lyase
KW - Enzyme function
KW - Itaconate catabolism
KW - Leucine catabolism
KW - Mycobacterium
KW - Tuberculosis
UR - http://www.scopus.com/inward/record.url?scp=85070207022&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85070207022&partnerID=8YFLogxK
U2 - 10.1073/pnas.1906606116
DO - 10.1073/pnas.1906606116
M3 - Article
C2 - 31320588
AN - SCOPUS:85070207022
VL - 116
SP - 15907
EP - 15913
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 32
ER -