title: In vivo biosynthesis of terpene nucleosides provides unique chemical markers of Mycobacterium tuberculosis infection.
abstract: Although small molecules shed from pathogens are widely used to diagnose infection, such tests have not been widely implemented for tuberculosis. Here we show that the recently identified compound, 1-tuberculosinyladenosine (1-TbAd), accumulates to comprise >1% of all Mycobacterium tuberculosis lipid. In vitro and in vivo, two isomers of TbAd were detected that might serve as infection markers. Using mass spectrometry and nuclear magnetic resonance, we established the structure of the previously unknown molecule, N(6)-tuberculosinyladenosine (N(6)-TbAd). Its biosynthesis involves enzymatic production of 1-TbAd by Rv3378c followed by conversion to N(6)-TbAd via the Dimroth rearrangement. Intact biosynthetic genes are observed only within M. tuberculosis complex bacteria, and TbAd was not detected among other medically important pathogens, environmental bacteria, and vaccine strains. With no substantially similar known molecules in nature, the discovery and in vivo detection of two abundant terpene nucleosides support their development as specific diagnostic markers of tuberculosis.
authors.affiliation: Division of Rheumatology, Immunology and Allergy, Brigham and Women's Hospital, Harvard Medical School, Smith Building Room 538, 1 Jimmy Fund Way, Boston, MA 02115, USA.
authors.forename: David C
authors.initials: DC
authors.lastname: Young
authors.affiliation: Division of Rheumatology, Immunology and Allergy, Brigham and Women's Hospital, Harvard Medical School, Smith Building Room 538, 1 Jimmy Fund Way, Boston, MA 02115, USA.
authors.forename: Emilie
authors.initials: E
authors.lastname: Layre
authors.affiliation: K-RITH, KwuZulu-Natal Research Institute for Tuberculosis & HIV, Nelson R. Mandela School of Medicine-University of Kwazulu-Natal, K-RITH Tower Building, 719 Umbilo Road, Durban, 4001 Private Bag X7, Congela-Durban 4001, South Africa.
authors.forename: Shih-Jung
authors.initials: SJ
authors.lastname: Pan
authors.affiliation: K-RITH, KwuZulu-Natal Research Institute for Tuberculosis & HIV, Nelson R. Mandela School of Medicine-University of Kwazulu-Natal, K-RITH Tower Building, 719 Umbilo Road, Durban, 4001 Private Bag X7, Congela-Durban 4001, South Africa; UCSF School of Medicine, San Francisco, CA 94143, USA.
authors.forename: Asa
authors.initials: A
authors.lastname: Tapley
authors.affiliation: K-RITH, KwuZulu-Natal Research Institute for Tuberculosis & HIV, Nelson R. Mandela School of Medicine-University of Kwazulu-Natal, K-RITH Tower Building, 719 Umbilo Road, Durban, 4001 Private Bag X7, Congela-Durban 4001, South Africa; Center for Tuberculosis Research, Division of Infections Diseases, Johns Hopkins University School of Medicine, 1550 Orleans Street Room 108, Baltimore, MD 21231, USA.
authors.forename: John
authors.initials: J
authors.lastname: Adamson
authors.affiliation: Division of Rheumatology, Immunology and Allergy, Brigham and Women's Hospital, Harvard Medical School, Smith Building Room 538, 1 Jimmy Fund Way, Boston, MA 02115, USA.
authors.forename: Chetan
authors.initials: C
authors.lastname: Seshadri
authors.affiliation: Stratingh Institute for Chemistry, University of Groningen, Nijenborgh 7, 9747 AG Groningen, The Netherlands.
authors.forename: Zhongtao
authors.initials: Z
authors.lastname: Wu
authors.affiliation: Stratingh Institute for Chemistry, University of Groningen, Nijenborgh 7, 9747 AG Groningen, The Netherlands.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Buter
authors.affiliation: Stratingh Institute for Chemistry, University of Groningen, Nijenborgh 7, 9747 AG Groningen, The Netherlands.
authors.forename: Adriaan J
authors.initials: AJ
authors.lastname: Minnaard
authors.affiliation: Swiss Tropical and Public Health Institute, 4051 Basel, Switzerland; University of Basel, 4003 Basel, Switzerland.
authors.forename: Mireia
authors.initials: M
authors.lastname: Coscolla
authors.affiliation: Swiss Tropical and Public Health Institute, 4051 Basel, Switzerland; University of Basel, 4003 Basel, Switzerland.
authors.forename: Sebastien
authors.initials: S
authors.lastname: Gagneux
authors.affiliation: Division of Infectious Diseases & Immunology, New York University School of Medicine, 522 First Avenue, SRB 901, New York, NY 10016, USA.
authors.forename: Richard
authors.initials: R
authors.lastname: Copin
authors.affiliation: Division of Infectious Diseases & Immunology, New York University School of Medicine, 522 First Avenue, SRB 901, New York, NY 10016, USA.
authors.forename: Joel D
authors.initials: JD
authors.lastname: Ernst
authors.affiliation: K-RITH, KwuZulu-Natal Research Institute for Tuberculosis & HIV, Nelson R. Mandela School of Medicine-University of Kwazulu-Natal, K-RITH Tower Building, 719 Umbilo Road, Durban, 4001 Private Bag X7, Congela-Durban 4001, South Africa; Center for Tuberculosis Research, Division of Infections Diseases, Johns Hopkins University School of Medicine, 1550 Orleans Street Room 108, Baltimore, MD 21231, USA.
authors.forename: William R
authors.initials: WR
authors.lastname: Bishai
authors.affiliation: Department of Chemistry MS015, Brandeis University, Waltham, MA 02453-2728, USA.
authors.forename: Barry B
authors.initials: BB
authors.lastname: Snider
authors.affiliation: Division of Rheumatology, Immunology and Allergy, Brigham and Women's Hospital, Harvard Medical School, Smith Building Room 538, 1 Jimmy Fund Way, Boston, MA 02115, USA. Electronic address: bmoody@partners.org.
authors.forename: D Branch
authors.initials: DB
authors.lastname: Moody
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D007536:Isomerism; D008055:Lipids / Q000032:analysis / Q000096:biosynthesis* / Q000302:isolation & purification; D008168:Lung / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008807:Mice, Inbred BALB C; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000378:metabolism*; D009705:Nucleosides / Q000032:analysis* / Q000096:biosynthesis / Q000737:chemistry; D020641:Polymorphism, Single Nucleotide; D021241:Spectrometry, Mass, Electrospray Ionization; D013729:Terpenes / Q000737:chemistry*; D014376:Tuberculosis / Q000175:diagnosis*
chemical_list: C587948:1-tuberculosinyladenosine; D001426:Bacterial Proteins; D008055:Lipids; D009705:Nucleosides; D013729:Terpenes