Intrinsic Folding Properties of the HLA-B27 Heavy Chain Revealed by Single Chain Trimer Versions of Peptide-Loaded Class I Major Histocompatibility Complex Molecules

Lenart, Izabela, Truong, Linh-Huyen, Nguyen, Dinh Dung, Rasiukienė, Olga, Tsao, Edward, Armstrong, Jonathan, Kumar, Pankaj, McHugh, Kirsty, Pereira, Branca I., Maan, Balraj S., Garstka, Malgorzata A., Bowness, Paul, Blake, Neil, Powis, Simon J., Gould, Keith, Nesbeth, Darren and Antoniou, Antony N. (2022) Intrinsic Folding Properties of the HLA-B27 Heavy Chain Revealed by Single Chain Trimer Versions of Peptide-Loaded Class I Major Histocompatibility Complex Molecules. Frontiers in Immunology, 13. p. 902135. ISSN 1664-3224

Preview

Text fimmu-13-902135.pdf - Published Version Available under License Creative Commons Attribution 4.0. Download (3MB) | Preview

Abstract

Peptide-loaded Major Histocompatibility Complex (pMHC) class I molecules can be expressed in a single chain trimeric (SCT) format, composed of a specific peptide fused to the light chain beta-2 microglobulin (β2m) and MHC class I heavy chain (HC) by flexible linker peptides. pMHC SCTs have been used as effective molecular tools to investigate cellular immunity and represent a promising vaccine platform technology, due to their intracellular folding and assembly which is apparently independent of host cell folding pathways and chaperones. However, certain MHC class I HC molecules, such as the Human Leukocyte Antigen B27 (HLA-B27) allele, present a challenge due to their tendency to form HC aggregates. We constructed a series of single chain trimeric molecules to determine the behaviour of the HLA-B27 HC in a scenario that usually allows for efficient MHC class I molecule folding. When stably expressed, a pMHC SCT incorporating HLA-B27 HC formed chaperone-bound homodimers within the endoplasmic reticulum (ER). A series of HLA-B27 SCT substitution mutations revealed that the F pocket and antigen binding groove regions of the HLA-B27 HC defined the folding and dimerisation of the single chain complex, independently of the peptide sequence. Furthermore, pMHC SCTs can demonstrate variability in their association with the intracellular antigen processing machinery.

Actions (login required)

View Item

Downloads