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J Biol Chem, Vol. 275, Issue 3, 1581-1586, January 21, 2000
From Tumor Immunology, Lund University, Solvegatan 21, s-22362 Lund, Sweden
The transporter associated with antigen
processing (TAP) binds peptides in its cytosolic part and subsequently
translocates the peptides into the lumen of the endoplasmic reticulum
(ER), where assembly of major histocompatibility complex (MHC) class I
and peptide takes place. Tapasin is a subunit of the TAP complex and
binds both to TAP1 and MHC class I. In the absence of tapasin, the
assembly of MHC class I in the ER is impaired, and the surface expression is reduced. To clarify the function of tapasin in the processing of antigenic peptides, we studied the interaction of peptide
and TAP, peptide transport across the membrane of the ER, and
association of peptides with MHC class I molecules in the microsomes
derived from tapasin mutant cell line 721.220, its sister cell line
721.221 expressing tapasin, and their HLA-A2 transfectants. The binding
of peptides to TAP in tapasin mutant 721.220 cells was significantly
diminished in comparison with 721.221 cells. Impaired peptide-TAP
interaction resulted in a defective peptide transport in tapasin mutant
721.220 cells. Interestingly, despite the diminished peptide binding to
TAP, the transport rate of TAP-associated peptides was not
significantly altered in 721.220 cells. After transfection of tapasin
cDNA into 721.220 cells, efficient peptide-TAP interaction was
restored. Thus, we conclude that tapasin is required for efficient
peptide-TAP interaction.
Tapasin Is Required for Efficient Peptide Binding to Transporter
Associated with Antigen Processing*
,
*
This study was supported by Crafoordska Stiftelsen Grant
990589, Swedish Cancer Society Grant 3975-B98-02XBB, a grant from the
Medical faculty, Lund University, and grants from the Foundation for
Strategic Research (Infection and Vaccionlogy Grant 36/98 and
Inflammation Research Program Grant 99).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed. Tel.: 46-46-2227848;
Fax: 46-46-2224606; E-mail: su-ling.li@wblab.lu.se and
ping.wang@wblab.lu.se.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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