Uptake of Unnatural Trehalose Analogs As a Reporter for Mycobacterium Tuberculosis

Overview

Journal Nat Chem Biol

Specialties Biochemistry
Biology
Chemistry

Date 2011 Mar 8

PMID 21378984

Citations 106

Authors

Keriann M Backus

Helena I Boshoff

Conor S Barry

Omar Boutureira

Mitul K Patel

Francois DHooge

Seung Seo Lee

Laura E Via

Kapil Tahlan

Clifton E Barry 3rd

Benjamin G Davis

Affiliations

Soon will be listed here.

Abstract

The detection of tuberculosis currently relies upon insensitive and unspecific techniques; newer diagnostics would ideally co-opt specific bacterial processes to provide real-time readouts. The trehalose mycolyltransesterase enzymes (antigens 85A, 85B and 85C (Ag85A, Ag85B, Ag85C)) serve as essential mediators of cell envelope function and biogenesis in Mycobacterium tuberculosis. Through the construction of a systematically varied sugar library, we show here that Ag85 enzymes have exceptionally broad substrate specificity. This allowed exogenously added synthetic probes to be specifically incorporated into M. tuberculosis growing in vitro and within macrophages. Even bulky substituents, such as a fluorescein-containing trehalose probe (FITC-trehalose), were incorporated by growing bacilli, thereby producing fluorescent bacteria; microscopy revealed selective labeling of poles and membrane. Addition of FITC-trehalose to M. tuberculosis-infected macrophages allowed selective, sensitive detection of M. tuberculosis within infected mammalian macrophages. These studies suggest that analogs of trehalose may prove useful as probes of function and for other imaging modalities.

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