Prominence of β 2 -Microglobulin, Class I Heavy Chain Conformation, and Tapasin in the Interactions of Class I Heavy Chain with Calreticulin and the Transporter Associated with Antigen Processing

Joyce C Solheim, Michael R. Harris, Cathy S. Kindle, Ted H. Hansen

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115 Citations (Scopus)

Abstract

Newly synthesized class I heavy (H) chain/β 2 m heterodimers awaiting peptides in the endoplasmic reticulum are associated with the transporter associated with Ag processing (TAP). We present evidence here that calreticulin, but not calnexin, displays steady state association with class I/TAP complexes. To separate the ability of β 2 m to bind with TAP and calreticulin from that of H chain, we studied human cell lines that lack expression of β 2 m or H chain. Little if any H chain was detected in association with TAP and calreticulin in the β 2 m - cell line Daudi. By contrast, high levels of β 2 m are found with TAP and calreticulin in the H chain-deficient cell line LCL 721.221, even after preclearance of the trace amount of class IB protein expressed by this cell line. Thus, β 2 m appears to bind TAP in the absence of H chain, providing an elegant mechanism to retain β 2 m in the endoplasmic reticulum at the site of peptide loading. To investigate whether other molecules participate in the binding of β 2 m and H chain to TAP and calreticulin, we analyzed the deletion mutant cell line LCL 721.220, which lacks tapasin. In 721.220, TAP and calreticulin are not associated with each other. Also, in these cells, H chain/β 2 m are not associated with TAP, but H chain and a low level of β 2 m are associated with calreticulin. These results suggest that tapasin is an obligatory mediator of the assemblage of calreticulin/H chain/β 2 m with TAP.

Original languageEnglish (US)
Pages (from-to)2236-2241
Number of pages6
JournalJournal of Immunology
Volume158
Issue number5
StatePublished - Mar 1 1997

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Calreticulin
Antigen Presentation
Cell Line
Endoplasmic Reticulum
Calnexin
Peptides
tapasin

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Prominence of β 2 -Microglobulin, Class I Heavy Chain Conformation, and Tapasin in the Interactions of Class I Heavy Chain with Calreticulin and the Transporter Associated with Antigen Processing . / Solheim, Joyce C; Harris, Michael R.; Kindle, Cathy S.; Hansen, Ted H.

In: Journal of Immunology, Vol. 158, No. 5, 01.03.1997, p. 2236-2241.

Research output: Contribution to journalArticle

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abstract = "Newly synthesized class I heavy (H) chain/β 2 m heterodimers awaiting peptides in the endoplasmic reticulum are associated with the transporter associated with Ag processing (TAP). We present evidence here that calreticulin, but not calnexin, displays steady state association with class I/TAP complexes. To separate the ability of β 2 m to bind with TAP and calreticulin from that of H chain, we studied human cell lines that lack expression of β 2 m or H chain. Little if any H chain was detected in association with TAP and calreticulin in the β 2 m - cell line Daudi. By contrast, high levels of β 2 m are found with TAP and calreticulin in the H chain-deficient cell line LCL 721.221, even after preclearance of the trace amount of class IB protein expressed by this cell line. Thus, β 2 m appears to bind TAP in the absence of H chain, providing an elegant mechanism to retain β 2 m in the endoplasmic reticulum at the site of peptide loading. To investigate whether other molecules participate in the binding of β 2 m and H chain to TAP and calreticulin, we analyzed the deletion mutant cell line LCL 721.220, which lacks tapasin. In 721.220, TAP and calreticulin are not associated with each other. Also, in these cells, H chain/β 2 m are not associated with TAP, but H chain and a low level of β 2 m are associated with calreticulin. These results suggest that tapasin is an obligatory mediator of the assemblage of calreticulin/H chain/β 2 m with TAP.",
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