BTN3A1 discriminates γδ T cell phosphoantigens from non-antigenic small molecules via a conformational sensor in its B30.2 domain

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BTN3A1 discriminates γδ T cell phosphoantigens from non-antigenic small molecules via a conformational sensor in its B30.2 domain. / Salim, Mahboob; Knowles, Timothy J; Baker, Alfie T; Davey, Martin S; Jeeves, Mark; Sridhar, Pooja; Wilkie, John; Willcox, Carrie R; Kadri, Hachemi; Taher, Taher E; Vantourout, Pierre; Hayday, Adrian; Mehellou, Youcef; Mohammed, Fiyaz; Willcox, Benjamin E.

In: ACS chemical biology, Vol. 12, No. 10, 14.09.2017, p. 2631-2643.

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@article{0f0a69792d414a29aff81f032d68324a,
title = "BTN3A1 discriminates γδ T cell phosphoantigens from non-antigenic small molecules via a conformational sensor in its B30.2 domain",
abstract = "Human Vγ9/Vδ2 T-cells detect tumour cells and microbial infections by recognising small phosphorylated prenyl metabolites termed phosphoantigens (P-Ag). The type-1 transmembrane protein Butyrophilin 3A1 (BTN3A1) is critical to the P-Ag-mediated activation of Vγ9/Vδ2 T-cells, however, the molecular mechanisms involved in BTN3A1-mediated metabolite sensing are unclear, including how P-Ag are discriminated from non-antigenic small molecules. Here, we utilised NMR and X-ray crystallography to probe P-Ag sensing by BTN3A1. Whereas the BTN3A1 Immunoglobulin Variable domain failed to bind P-Ag, the intracellular B30.2 domain bound a range of negatively-charged small molecules, including P-Ag, in a positively-charged surface pocket. However, NMR chemical shift perturbations indicated BTN3A1 discriminated P-Ag from non-antigenic small molecules by their ability to induce a specific conformational change in the B30.2 domain that propagated from the P-Ag binding site to distal parts of the domain. These results suggest BTN3A1 selectively detects P-Ag intracellularly via a conformational antigenic sensor in its B30.2 domain, and have implications for rational design of antigens for Vγ9/Vδ2 -based T-cell immunotherapies. ",
keywords = "Antigens, Antigens, CD/genetics, Butyrophilins/genetics, Cloning, Molecular, Coculture Techniques, Gene Expression Regulation/physiology, HEK293 Cells, Humans, Magnetic Resonance Spectroscopy, Models, Chemical, Mutation, Phosphoproteins, Protein Conformation, Protein Domains, Receptors, Antigen, T-Cell, gamma-delta/genetics, T-Lymphocytes/metabolism",
author = "Mahboob Salim and Knowles, {Timothy J} and Baker, {Alfie T} and Davey, {Martin S} and Mark Jeeves and Pooja Sridhar and John Wilkie and Willcox, {Carrie R} and Hachemi Kadri and Taher, {Taher E} and Pierre Vantourout and Adrian Hayday and Youcef Mehellou and Fiyaz Mohammed and Willcox, {Benjamin E}",
year = "2017",
month = sep
day = "14",
doi = "10.1021/acschembio.7b00694",
language = "English",
volume = "12",
pages = "2631--2643",
journal = "ACS chemical biology",
issn = "1554-8929",
publisher = "American Chemical Society",
number = "10",

}

RIS

TY - JOUR

T1 - BTN3A1 discriminates γδ T cell phosphoantigens from non-antigenic small molecules via a conformational sensor in its B30.2 domain

AU - Salim, Mahboob

AU - Knowles, Timothy J

AU - Baker, Alfie T

AU - Davey, Martin S

AU - Jeeves, Mark

AU - Sridhar, Pooja

AU - Wilkie, John

AU - Willcox, Carrie R

AU - Kadri, Hachemi

AU - Taher, Taher E

AU - Vantourout, Pierre

AU - Hayday, Adrian

AU - Mehellou, Youcef

AU - Mohammed, Fiyaz

AU - Willcox, Benjamin E

PY - 2017/9/14

Y1 - 2017/9/14

N2 - Human Vγ9/Vδ2 T-cells detect tumour cells and microbial infections by recognising small phosphorylated prenyl metabolites termed phosphoantigens (P-Ag). The type-1 transmembrane protein Butyrophilin 3A1 (BTN3A1) is critical to the P-Ag-mediated activation of Vγ9/Vδ2 T-cells, however, the molecular mechanisms involved in BTN3A1-mediated metabolite sensing are unclear, including how P-Ag are discriminated from non-antigenic small molecules. Here, we utilised NMR and X-ray crystallography to probe P-Ag sensing by BTN3A1. Whereas the BTN3A1 Immunoglobulin Variable domain failed to bind P-Ag, the intracellular B30.2 domain bound a range of negatively-charged small molecules, including P-Ag, in a positively-charged surface pocket. However, NMR chemical shift perturbations indicated BTN3A1 discriminated P-Ag from non-antigenic small molecules by their ability to induce a specific conformational change in the B30.2 domain that propagated from the P-Ag binding site to distal parts of the domain. These results suggest BTN3A1 selectively detects P-Ag intracellularly via a conformational antigenic sensor in its B30.2 domain, and have implications for rational design of antigens for Vγ9/Vδ2 -based T-cell immunotherapies.

AB - Human Vγ9/Vδ2 T-cells detect tumour cells and microbial infections by recognising small phosphorylated prenyl metabolites termed phosphoantigens (P-Ag). The type-1 transmembrane protein Butyrophilin 3A1 (BTN3A1) is critical to the P-Ag-mediated activation of Vγ9/Vδ2 T-cells, however, the molecular mechanisms involved in BTN3A1-mediated metabolite sensing are unclear, including how P-Ag are discriminated from non-antigenic small molecules. Here, we utilised NMR and X-ray crystallography to probe P-Ag sensing by BTN3A1. Whereas the BTN3A1 Immunoglobulin Variable domain failed to bind P-Ag, the intracellular B30.2 domain bound a range of negatively-charged small molecules, including P-Ag, in a positively-charged surface pocket. However, NMR chemical shift perturbations indicated BTN3A1 discriminated P-Ag from non-antigenic small molecules by their ability to induce a specific conformational change in the B30.2 domain that propagated from the P-Ag binding site to distal parts of the domain. These results suggest BTN3A1 selectively detects P-Ag intracellularly via a conformational antigenic sensor in its B30.2 domain, and have implications for rational design of antigens for Vγ9/Vδ2 -based T-cell immunotherapies.

KW - Antigens

KW - Antigens, CD/genetics

KW - Butyrophilins/genetics

KW - Cloning, Molecular

KW - Coculture Techniques

KW - Gene Expression Regulation/physiology

KW - HEK293 Cells

KW - Humans

KW - Magnetic Resonance Spectroscopy

KW - Models, Chemical

KW - Mutation

KW - Phosphoproteins

KW - Protein Conformation

KW - Protein Domains

KW - Receptors, Antigen, T-Cell, gamma-delta/genetics

KW - T-Lymphocytes/metabolism

U2 - 10.1021/acschembio.7b00694

DO - 10.1021/acschembio.7b00694

M3 - Article

C2 - 28862425

VL - 12

SP - 2631

EP - 2643

JO - ACS chemical biology

JF - ACS chemical biology

SN - 1554-8929

IS - 10

ER -