Calreticulin promotes folding of functional human leukocyte antigen class I molecules in vitro

Slobodan Culina, Gregoire Lauvau, Brigitte Gubler, Peter M. Van Endert

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

The assembly of MHC class I molecules with β2-microglobulin and peptides is assisted by the housekeeping chaperones calnexin, calreticulin, and Erp57 and the dedicated accessory protein, tapasin. Tapasin and calreticulin are essential for efficient MHC class I assembly, but their precise action during class I assembly remains to be elucidated. Previous in vitro studies have demonstrated that the lectin calreticulin interacts with monoglucosylated MHC class I heavy chains, whatever their state of assembly with light chains and peptide, and inhibits their aggregation above physiological temperature. We used a soluble single chain HLA-A2/β2- microglobulin molecule, A2SC, to study the effect of calreticulin on the peptide binding capacity of HLA class I molecules. Calreticulin inhibited the formation of A2SC aggregates both when co-expressed in insect cells and during incubations at elevated temperature. Calreticulin dramatically enhanced acquisition of peptide binding capacity when added to denatured A2SC molecules during refolding at 4 °C. However, it had no effect on the rapid loss of A2SC peptide binding capacity at physiological temperature. We conclude that calreticulin promotes the folding of HLA class I molecules to a state in which, at low temperature, they spontaneously acquire peptide binding capacity. However, it does not induce or maintain a peptide-receptive state of the class I-binding site, which is likely to be promoted by one or several other components of the class I loading complexes. By being amenable to complementation with additional proteins, the described system should be useful for identification of these components.

Original languageEnglish (US)
Pages (from-to)54210-54215
Number of pages6
JournalJournal of Biological Chemistry
Volume279
Issue number52
DOIs
StatePublished - Dec 24 2004
Externally publishedYes

Fingerprint

Calreticulin
HLA Antigens
Peptides
Molecules
Temperature
Calnexin
HLA-A2 Antigen
Housekeeping
Accessories
In Vitro Techniques
Lectins
Insects
Proteins
Agglomeration
Binding Sites
Light

ASJC Scopus subject areas

  • Biochemistry

Cite this

Calreticulin promotes folding of functional human leukocyte antigen class I molecules in vitro. / Culina, Slobodan; Lauvau, Gregoire; Gubler, Brigitte; Van Endert, Peter M.

In: Journal of Biological Chemistry, Vol. 279, No. 52, 24.12.2004, p. 54210-54215.

Research output: Contribution to journalArticle

Culina, Slobodan ; Lauvau, Gregoire ; Gubler, Brigitte ; Van Endert, Peter M. / Calreticulin promotes folding of functional human leukocyte antigen class I molecules in vitro. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 52. pp. 54210-54215.
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