Structure of the Mycobacterium tuberculosis proteasome and mechanism of inhibition by a peptidyl boronate

• Published in: Molecular microbiology 2006-03, Vol.59 (5), p.1417-1428
• Main Authors: Hu, Guiqing, Lin, Gang, Wang, Ming, Dick, Lawrence, Xu, Rui‐Ming, Nathan, Carl, Li, Huilin
• Format: Article
• Language: English
• Subjects: Bacteria; Binding Sites; Biological and medical sciences; Boronic Acids - chemistry; Boronic Acids - pharmacology; Bortezomib; Crystal structure; Crystallography, X-Ray; Dipeptides - chemistry; Dipeptides - pharmacology; Eubacterium; Fundamental and applied biological sciences. Psychology; Microbiology; Molecular Structure; Mycobacterium tuberculosis; Mycobacterium tuberculosis - enzymology; Peptides; Protease Inhibitors - chemistry; Proteases; Proteasome Endopeptidase Complex - chemistry; Proteasome Endopeptidase Complex - metabolism; Proteasome Inhibitors; Protein Subunits; Pyrazines - chemistry; Substrate Specificity; Tuberculosis
• ISSN: 0950-382X; 1365-2958
• DOI: 10.1111/j.1365-2958.2005.05036.x

Mycobacterium tuberculosis (Mtb) has the remarkable ability to resist killing by human macrophages. The 750 kDa proteasome, not available in most eubacteria except Actinomycetes, appears to contribute to Mtb's resistance. The crystal structure of the Mtb proteasome at 3.0 Å resolution reveals a substrate‐binding pocket with composite features of the distinct β1, β2 and β5 substrate binding sites of eukaryotic proteasomes, accounting for the broad specificity of the Mtb proteasome towards oligopeptides described in the companion article [Lin et al. (2006), Mol Microbiol doi:10.1111/j.1365‐2958.2005.05035.x]. The substrate entrance at the end of the cylindrical proteasome appears open in the crystal structure due to partial disorder of the α‐subunit N‐terminal residues. However, cryo‐electron microscopy of the core particle reveals a closed end, compatible with the density observed in negative‐staining electron microscopy that depended on the presence of the N‐terminal octapetides of the α‐subunits in the companion article, suggesting that the Mtb proteasome has a gated structure. We determine for the first time the proteasomal inhibition mechanism of the dipeptidyl boronate N‐(4‐morpholine)carbonyl‐β‐(1‐naphthyl)‐l‐alanine‐l‐leucine boronic acid (MLN‐273), an analogue of the antimyeloma drug bortezomib. The structure improves prospects for designing Mtb‐specific proteasomal inhibitors as a novel approach to chemotherapy of tuberculosis.