Uncovering Structural and Molecular Dynamics of ESAT-6:β2M Interaction: Asp53 of Human β2-Microglobulin Is Critical for the ESAT-6:β2M Complexation

Key Points Asp53 residue of β2M is important in forming a complex with Met93 of ESAT-6. SM09 and SM15 rescued ESAT-6–mediated downregulation of MHC class I Ag presentation. SM09 and SM15 inhibited survival of M. tuberculosis inside the macrophage. ESAT-6 is a small secreted protein of Mycobacterium tuberculosis involved in the ESAT-6 secretion system (ESX-1)–mediated virulence and pathogenesis. The protein interacts with β2M, causing downregulation of MHC class I Ag presentation, which could be one of the mechanisms by which it favors increased survival of the bacilli inside the host. In an earlier study, we have shown that the C-terminal region of ESAT-6 is crucial for its interaction with β2M. However, the interface of β2M involved in interaction with ESAT-6 and detailed physicochemical changes associated with ESAT-6:β2M complexation are not fully defined. In this study, using computational and site-directed mutagenesis studies, we demonstrate the presence of strong noncovalent hydrophobic interactions between ESAT-6 and β2M in addition to the vital hydrogen bonding between the aspartate residue (Asp53) of β2M and methionine (Met93) of ESAT-6. Docking-based high-throughput virtual screening followed by 16-point screening on microscale thermophoresis resulted in the identification of two potent inhibitors (SM09 and SM15) that mask the critical Met93 residue of ESAT-6 that is required for ESAT-6:β2M interaction and could rescue cell surface expression of β2M and HLA in human macrophages as well as MHC class I Ag presentation suppressed by ESAT-6 in peritoneal macrophages isolated from C57BL/6 mice. Both SM09 and SM15 significantly inhibited intracellular survival of M. tuberculosis in human macrophages. Further, we characterized the physicochemical properties involved in the ESAT-6:β2M complexation, which may help in understanding host–pathogen interactions.

• Publication year

  2019

• Venue

  Journal of Immunology

• Publication date

  2019-10-01

• Fields of study

  Medicine, Chemistry

• Identifiers
  DOI 10.4049/jimmunol.1700525PMID 31484733
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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