title: Total Synthesis of Mycobacterium tuberculosis Dideoxymycobactin-838 and Stereoisomers: Diverse CD1a-Restricted T Cells Display a Common Hierarchy of Lipopeptide Recognition.
abstract: Mycobacterium tuberculosis produces dideoxymycobactin-838 (DDM-838), a lipopeptide that potently activates T cells upon binding to the MHC-like antigen-presenting molecule CD1a. M. tuberculosis produces DDM-838 in only trace amounts and a previous solid-phase synthesis provided sub-milligram quantities. We describe a high-yielding solution-phase synthesis of DDM-838 that features a Mitsunobu substitution that avoids yield-limiting epimerization at lysine during esterification, and amidation conditions that prevent double-bond isomerization of the Z-C20:1 acyl chain, and provides material with equivalent antigenicity to natural DDM-838. Isomers of DDM-838 that varied in stereochemistry at the central lysine and the C20:1 acyl chain were compared for their ability to be recognised by CD1a-restricted T cell receptors (TCRs). These TCRs, derived from unrelated human donors, exhibited a similar spectrum of reactivity towards the panel of DDM-838 isomers, highlighting the exquisite sensitivity of lipopeptide-reactive T cells for the natural DDM stereochemistry.
authors.affiliation: School of Chemistry and Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, Victoria, 3010, Australia.|Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity, University of Melbourne, Parkville, Victoria, 3010, Australia.|Infection and Immunity Program, Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University, Clayton, VIC, 3800, Australia.|Australian Research Council Centre of Excellence for Advanced Molecular Imaging, University of Melbourne, Parkville, Victoria, 3010, Australia.
authors.forename: Janice M H
authors.initials: JM
authors.lastname: Cheng
authors.affiliation: Division of Chemistry and Structural Biology, Institute for Molecular Bioscience, The University of Queensland, Brisbane, Queensland, 4072, Australia.|Australian Research Council Centre of Excellence for Advanced Molecular Imaging, The University of Queensland, Brisbane, Queensland, 4072, Australia.
authors.forename: Ligong
authors.initials: L
authors.lastname: Liu
authors.affiliation: Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity, University of Melbourne, Parkville, Victoria, 3010, Australia.|Australian Research Council Centre of Excellence for Advanced Molecular Imaging, University of Melbourne, Parkville, Victoria, 3010, Australia.
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Pellicci
authors.affiliation: Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity, University of Melbourne, Parkville, Victoria, 3010, Australia.|Australian Research Council Centre of Excellence for Advanced Molecular Imaging, University of Melbourne, Parkville, Victoria, 3010, Australia.
authors.forename: Scott J J
authors.initials: SJ
authors.lastname: Reddiex
authors.affiliation: Division of Rheumatology, Immunology and Allergy, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, 02115, USA.
authors.forename: Rachel N
authors.initials: RN
authors.lastname: Cotton
authors.affiliation: Division of Rheumatology, Immunology and Allergy, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, 02115, USA.
authors.forename: Tan-Yun
authors.initials: TY
authors.lastname: Cheng
authors.affiliation: Division of Rheumatology, Immunology and Allergy, Brigham and Women's Hospital, Harvard Medical School, 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, Boston, MA, 02115, USA.|Department of Infectious Diseases and Immunology, School of Veterinary Medicine, University of Utrecht, The Netherlands.
authors.forename: Ildiko
authors.initials: I
authors.lastname: Van Rhijn
authors.affiliation: Division of Rheumatology, Immunology and Allergy, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, 02115, USA.
authors.forename: D Branch
authors.initials: DB
authors.lastname: Moody
authors.affiliation: Infection and Immunity Program, Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University, Clayton, VIC, 3800, Australia.|Australian Research Council Centre of Excellence for Advanced Molecular Imaging, Monash University, Clayton, Victoria, 3800, Australia.|Institute of Infection and Immunity, Cardiff University School of Medicine, Cardiff, CF14 4XN, UK.
authors.forename: Jamie
authors.initials: J
authors.lastname: Rossjohn
authors.affiliation: Division of Chemistry and Structural Biology, Institute for Molecular Bioscience, The University of Queensland, Brisbane, Queensland, 4072, Australia.|Australian Research Council Centre of Excellence for Advanced Molecular Imaging, The University of Queensland, Brisbane, Queensland, 4072, Australia.
authors.forename: David P
authors.initials: DP
authors.lastname: Fairlie
authors.affiliation: Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity, University of Melbourne, Parkville, Victoria, 3010, Australia.|Australian Research Council Centre of Excellence for Advanced Molecular Imaging, University of Melbourne, Parkville, Victoria, 3010, Australia.
authors.forename: Dale I
authors.initials: DI
authors.lastname: Godfrey
authors.affiliation: School of Chemistry and Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, Victoria, 3010, Australia.|Australian Research Council Centre of Excellence for Advanced Molecular Imaging, University of Melbourne, Parkville, Victoria, 3010, Australia.
authors.forename: Spencer J
authors.identifier: http://orcid.org/0000-0001-6341-4364
authors.initials: SJ
authors.lastname: Williams
mesh_terms: D018949:Antigens, CD1 / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D019169:Jurkat Cells; D055666:Lipopeptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D009169:Mycobacterium tuberculosis / Q000378:metabolism*; D010080:Oxazoles / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D013237:Stereoisomerism; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism*
chemical_list: D018949:Antigens, CD1; C419393:CD1a antigen; D055666:Lipopeptides; D010080:Oxazoles; D011994:Recombinant Proteins; C585579:dideoxymycobactin; D007371:Interferon-gamma
keywords: T cells; antigens; immunology; natural products; peptidolipids