TY - JOUR
T1 - TAP associates with a unique class I conformation, whereas calnexin associates with multiple class I forms in mouse and man
AU - Carreno, B. M.
AU - Solheim, J. C.
AU - Harris, M.
AU - Stroynowski, I.
AU - Connolly, J. M.
AU - Hansen, T. H.
PY - 1995
Y1 - 1995
N2 - To define the rules governing de novo assembly of the trimeric class I complex, we have identified the class I folding/assembly intermediates associated with calnexin or TAP, using both human and mouse cell lines. To better characterize the class I H chain structure associated with TAP, mouse mAb that distinguish open (64-3-7+) vs folded (30-5-7+) L(d) heavy (H) chains were used. We report here that open forms of L(d) are uniquely and specifically associated with TAP and that the conformational change in the class I H chain coincident with peptide binding induces TAP release. Chimeric L(d)/Q10 displayed TAP association, demonstrating that soluble class I molecules can bind TAP. As previously reported, β2m was found to be required for H chain association with TAP. Interestingly, β2m was associated with TAP in the human class I-negative cell line LCL 721.221, suggesting that β2m can bind to TAP before class I H chain. In contrast to TAP, which binds a specific class I conformation, calnexin was detected in association with multiple forms of both mouse and human class I. Most significantly, we show for the first time that β2m-assembled forms of human as well as mouse class I molecules interact with calnexin. Based on these findings, we propose a model for the sequential assembly of class I heterotrimers and their respective interactions with TAP and calnexin.
AB - To define the rules governing de novo assembly of the trimeric class I complex, we have identified the class I folding/assembly intermediates associated with calnexin or TAP, using both human and mouse cell lines. To better characterize the class I H chain structure associated with TAP, mouse mAb that distinguish open (64-3-7+) vs folded (30-5-7+) L(d) heavy (H) chains were used. We report here that open forms of L(d) are uniquely and specifically associated with TAP and that the conformational change in the class I H chain coincident with peptide binding induces TAP release. Chimeric L(d)/Q10 displayed TAP association, demonstrating that soluble class I molecules can bind TAP. As previously reported, β2m was found to be required for H chain association with TAP. Interestingly, β2m was associated with TAP in the human class I-negative cell line LCL 721.221, suggesting that β2m can bind to TAP before class I H chain. In contrast to TAP, which binds a specific class I conformation, calnexin was detected in association with multiple forms of both mouse and human class I. Most significantly, we show for the first time that β2m-assembled forms of human as well as mouse class I molecules interact with calnexin. Based on these findings, we propose a model for the sequential assembly of class I heterotrimers and their respective interactions with TAP and calnexin.
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M3 - Article
C2 - 7594473
AN - SCOPUS:0028858641
SN - 0022-1767
VL - 155
SP - 4726
EP - 4733
JO - Journal of Immunology
JF - Journal of Immunology
IS - 10
ER -