International Immunology, Vol 10, 1159-1165, Copyright © 1998 by Oxford University Press
GM Barton and AY Rudensky
We report an experimental system for abundant expression of specific
peptide-class II complexes in vivo and in vitro. We have constructed a
cassette which allows for the replacement of the CLIP region of invariant
chain (Ii) with an antigenic peptide. In fibroblasts expressing an altered
Ii protein, in which CLIP has been replaced with peptide 52-68 from the
class II I-E alpha chain (pEalpha), pEalpha-I-Ab complexes are formed with
high efficiency. This peptide loading occurs in the endoplasmic reticulum
(ER) when the Ii:pEalpha fusion protein associates with the I-Ab alpha and
beta chains. The trimeric complexes of Ii:pEalpha and I-Ab molecules are
stable in SDS and can be detected by the pEalpha-I-Ab-specific mAb, YAe,
indicating that pEalpha is bound in the class II groove in the context of
full-length Ii. These data strongly suggest that the CLIP region of intact
Ii prevents peptide loading in the ER by binding in the peptide binding
groove of newly synthesized class II alphabeta dimers.
ARTICLES
An altered invariant chain protein with an antigenic peptide in place of CLIP forms SDS-stable complexes with class II alphabeta dimers and facilitates highly efficient peptide loading
Department of Immunology, University of Washington School of Medicine, Seattle 98195, USA.
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