Issue 3, 2021

Bioorthogonal protein labelling enables the study of antigen processing of citrullinated and carbamylated auto-antigens

Abstract

Proteolysis is fundamental to many biological processes. In the immune system, it underpins the activation of the adaptive immune response: degradation of antigenic material into short peptides and presentation thereof on major histocompatibility complexes, leads to activation of T-cells. This initiates the adaptive immune response against many pathogens. Studying proteolysis is difficult, as the oft-used polypeptide reporters are susceptible to proteolytic sequestration themselves. Here we present a new approach that allows the imaging of antigen proteolysis throughout the processing pathway in an unbiased manner. By incorporating bioorthogonal functionalities into the protein in place of methionines, antigens can be followed during degradation, whilst leaving reactive sidechains open to templated and non-templated post-translational modifications, such as citrullination and carbamylation. Using this approach, we followed and imaged the post-uptake fate of the commonly used antigen ovalbumin, as well as the post-translationally citrullinated and/or carbamylated auto-antigen vinculin in rheumatoid arthritis, revealing differences in antigen processing and presentation.

Graphical abstract: Bioorthogonal protein labelling enables the study of antigen processing of citrullinated and carbamylated auto-antigens

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Article information

Article type
Paper
Submitted
13 1 2021
Accepted
22 2 2021
First published
23 2 2021
This article is Open Access
Creative Commons BY-NC license

RSC Chem. Biol., 2021,2, 855-862

Bioorthogonal protein labelling enables the study of antigen processing of citrullinated and carbamylated auto-antigens

T. van Leeuwen, C. Araman, L. Pieper Pournara, A. S. B. Kampstra, T. Bakkum, M. H. S. Marqvorsen, C. R. Nascimento, G. J. M. Groenewold, W. van der Wulp, M. G. M. Camps, G. M. C. Janssen, P. A. van Veelen, G. J. P. van Westen, A. P. A. Janssen, B. I. Florea, H. S. Overkleeft, F. A. Ossendorp, R. E. M. Toes and S. I. van Kasteren, RSC Chem. Biol., 2021, 2, 855 DOI: 10.1039/D1CB00009H

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