TY - JOUR
T1 - Azido Inositol Probes Enable Metabolic Labeling of Inositol-Containing Glycans and Reveal an Inositol Importer in Mycobacteria
AU - Hodges, Heather
AU - Obeng, Kwaku
AU - Avanzi, Charlotte
AU - Ausmus, Alex P.
AU - Angala, Shiva Kumar
AU - Kalera, Karishma
AU - Palcekova, Zuzana
AU - Swarts, Benjamin M.
AU - Jackson, Mary
N1 - Funding Information:
This work was supported by the National Institute of Allergy and Infectious Diseases (NIAID)/National Institutes of Health (NIH) grants AI064798 and AI155674 (M.J.), National Institute of General Medical Sciences (NIGMS)/National Institutes of Health (NIH) grant GM133080 (H.L.H.), NSF CAREER Award 1654408 (B.M.S.), and Camille and Henry Dreyfus Foundation Henry Dreyfus Teacher–Scholar Award TH-17-034 (B.M.S.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. NMR instrumentation at Central Michigan University was supported by NSF MRI Award 2117338 (B.M.S.).
Publisher Copyright:
© 2023 American Chemical Society.
PY - 2023/3/17
Y1 - 2023/3/17
N2 - Bacteria from the genus Mycobacterium include pathogens that cause serious diseases in humans and remain as difficult infectious agents to treat. Central to these challenges are the composition and organization of the mycobacterial cell envelope, which includes unique and complex glycans. Inositol is an essential metabolite for mycobacteria due to its presence in the structural core of the immunomodulatory cell envelope glycolipids phosphatidylinositol mannoside (PIM) and PIM-anchored lipomannan (LM) and lipoarabinomannan (LAM). Despite their importance to mycobacterial physiology and pathogenesis, many aspects of PIM, LM, and LAM construction and dynamics remain poorly understood. Recently, probes that allow metabolic labeling and detection of specific mycobacterial glycans have been developed to investigate cell envelope assembly and dynamics. However, these tools have been limited to peptidoglycan, arabinogalactan, and mycolic acid-containing glycolipids. Herein, we report the development of synthetic azido inositol (InoAz) analogues as probes that can metabolically label PIMs, LM, and LAM in intact mycobacteria. Additionally, we leverage an InoAz probe to discover an inositol importer and catabolic pathway in Mycobacterium smegmatis. We anticipate that in the future, InoAz probes, in combination with bioorthogonal chemistry, will provide a valuable tool for investigating PIM, LM, and LAM biosynthesis, transport, and dynamics in diverse mycobacterial organisms.
AB - Bacteria from the genus Mycobacterium include pathogens that cause serious diseases in humans and remain as difficult infectious agents to treat. Central to these challenges are the composition and organization of the mycobacterial cell envelope, which includes unique and complex glycans. Inositol is an essential metabolite for mycobacteria due to its presence in the structural core of the immunomodulatory cell envelope glycolipids phosphatidylinositol mannoside (PIM) and PIM-anchored lipomannan (LM) and lipoarabinomannan (LAM). Despite their importance to mycobacterial physiology and pathogenesis, many aspects of PIM, LM, and LAM construction and dynamics remain poorly understood. Recently, probes that allow metabolic labeling and detection of specific mycobacterial glycans have been developed to investigate cell envelope assembly and dynamics. However, these tools have been limited to peptidoglycan, arabinogalactan, and mycolic acid-containing glycolipids. Herein, we report the development of synthetic azido inositol (InoAz) analogues as probes that can metabolically label PIMs, LM, and LAM in intact mycobacteria. Additionally, we leverage an InoAz probe to discover an inositol importer and catabolic pathway in Mycobacterium smegmatis. We anticipate that in the future, InoAz probes, in combination with bioorthogonal chemistry, will provide a valuable tool for investigating PIM, LM, and LAM biosynthesis, transport, and dynamics in diverse mycobacterial organisms.
UR - http://www.scopus.com/inward/record.url?scp=85149477440&partnerID=8YFLogxK
U2 - 10.1021/acschembio.2c00912
DO - 10.1021/acschembio.2c00912
M3 - Article
C2 - 36856664
AN - SCOPUS:85149477440
SN - 1554-8929
VL - 18
SP - 595
EP - 604
JO - ACS Chemical Biology
JF - ACS Chemical Biology
IS - 3
ER -