Protein purification and crystallization of HLA-A∗02:01 in complex with SARS-CoV-2 peptides

Demetra S M Chatzileontiadou; Christopher Szeto; Dhilshan Jayasinghe; Stephanie Gras

doi:10.1016/j.xpro.2021.100635

Protein purification and crystallization of HLA-A∗02:01 in complex with SARS-CoV-2 peptides

STAR Protoc. 2021 Sep 17;2(3):100635. doi: 10.1016/j.xpro.2021.100635. Epub 2021 Jun 9.

Authors

Demetra S M Chatzileontiadou^{1

2}, Christopher Szeto^{1

2}, Dhilshan Jayasinghe^{1

2}, Stephanie Gras^{1

2}

Affiliations

¹ Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Bundoora, VIC 3083, Australia.
² Department of Biochemistry and Molecular Biology, Monash University, Clayton, VIC 3800, Australia.

Abstract

Understanding T-cell responses requires identifying viral peptides presented by human leukocyte antigens (HLAs). X-ray crystallography can be used to visualize their presentation. This protocol describes the expression, purification, and crystallization of HLA-A∗02:01, one of the most frequent HLA in the global population in complex with peptides derived from the SARS-CoV-2 nucleocapsid protein. This protocol can be applied to different HLA class I molecules bound to other peptides. For complete details on the use and execution of this protocol, please refer to Szeto et al. (2021).

Keywords: Immunology; Protein expression and purification; X-ray Crystallography.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

COVID-19 / immunology
COVID-19 / metabolism*
COVID-19 / virology
Coronavirus Nucleocapsid Proteins / chemistry*
Coronavirus Nucleocapsid Proteins / isolation & purification
Coronavirus Nucleocapsid Proteins / metabolism
Crystallography, X-Ray
Epitopes, T-Lymphocyte / immunology
HLA-A2 Antigen / chemistry*
HLA-A2 Antigen / metabolism
Humans
Peptide Fragments / chemistry*
Peptide Fragments / isolation & purification
Peptide Fragments / metabolism
Phosphoproteins / chemistry
Phosphoproteins / isolation & purification
Phosphoproteins / metabolism
SARS-CoV-2 / metabolism*
T-Lymphocytes / immunology*

Substances

Coronavirus Nucleocapsid Proteins
Epitopes, T-Lymphocyte
HLA-A*02:01 antigen
HLA-A2 Antigen
Peptide Fragments
Phosphoproteins
nucleocapsid phosphoprotein, SARS-CoV-2