Azido Inositol Probes Enable Metabolic Labeling of Inositol-Containing Glycans and Reveal an Inositol Importer in Mycobacteria

Hodges, Heather L.; Obeng, Kwaku; Avanzi, Charlotte; Ausmus, Alex P.; Angala, Shiva K.; Kalera, Karishma; Palčeková, Zuzana; Swarts, Benjamin M.; Jackson, Mary

doi:10.1021/acschembio.2c00912

Cited by 3 publications

(2 citation statements)

References 59 publications

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“…The only known probe of the PIMs, LM, and LAM is based on the metabolic incorporation of azide-functionalized inositol . Competition with the endogenous substrate occurs in multiple steps, so selective probe incorporation is contingent upon deleting genes in the endogenous inositol biosynthetic pathway.…”

Section: Introductionmentioning

confidence: 99%

“…21,24,26−32 The only known probe of the PIMs, LM, and LAM is based on the metabolic incorporation of azide-functionalized inositol. 33 Competition with the endogenous substrate occurs in multiple steps, so selective probe incorporation is contingent upon deleting genes in the endogenous inositol biosynthetic pathway. Though this requirement prevents the use of this probe in wild-type cells, these metabolic incorporation probes led to the discovery of a previously unidentified inositol transporter.…”

Section: ■ Introductionmentioning

confidence: 99%

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Selective Glycan Labeling of Mannose-Containing Glycolipids in Mycobacteria

Lee,

Marando,

Smelyansky

et al. 2023

J. Am. Chem. Soc.

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Mycobacterium tuberculosis (Mtb) is one of history's most successful human pathogens. By subverting typical immune responses, Mtb can persist within a host until conditions become favorable for growth and proliferation. Virulence factors that enable mycobacteria to modulate host immune systems include a suite of mannose-containing glycolipids: phosphatidylinositol mannosides, lipomannan, and lipoarabinomannan (LAM). Despite their importance, tools for their covalent capture, modification, and imaging are limited. Here, we describe a chemical biology strategy to detect and visualize these glycans. Our approach, biosynthetic incorporation, is to synthesize a lipid-glycan precursor that can be incorporated at a latestage step in glycolipid biosynthesis. We previously demonstrated selective mycobacterial arabinan modification by biosynthetic incorporation using an exogenous donor. This report reveals that biosynthetic labeling is general and selective: it allows for cell surface mannose-containing glycolipid modification without nonspecific labeling of mannosylated glycoproteins. Specifically, we employed azido-(Z,Z)-farnesyl phosphoryl-β-D-mannose probes and took advantage of the strain-promoted azide−alkyne cycloaddition to label and directly visualize the localization and dynamics of mycobacterial mannose-containing glycolipids. Our studies highlight the generality and utility of biosynthetic incorporation as the probe structure directs the selective labeling of distinct glycans. The disclosed agents allowed for direct tracking of the target immunomodulatory glycolipid dynamics in cellulo. We anticipate that these probes will facilitate investigating the diverse biological roles of these glycans.

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Section: Introductionmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%