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. 1986 Feb;83(3):747–751. doi: 10.1073/pnas.83.3.747

Identification of functional regions on the I-Ab molecule by site-directed mutagenesis.

L E Cohn, L H Glimcher, R A Waldmann, J A Smith, A Ben-Nun, J G Seidman, E Choi
PMCID: PMC322942  PMID: 2418441

Abstract

Functional analysis of mutant class II major histocompatibility complex molecules has begun to identify regions important for antibody binding and for T-cell activation. By using in vitro mutagenesis directed at the beta 1 domain of the Ab beta gene we have constructed three structurally distinct mutant Ab beta genes. Each of these genes, as well as the wild-type Ab beta gene, was cotransfected together with the wild-type Ab alpha gene into the Ia-negative B-lymphoma cell line M12.C3. Transfection resulted in the successful synthesis and cell surface expression of three mutant class II antigens that showed serological and functional alterations as compared to the I-Ab antigens from the M12.C3 cell transfected with the wild-type gene. The variable patterns of both I-Ab-specific monoclonal antibody binding and activation of I-Ab-specific T-cell hybridomas show that the mutations result in the loss of structural epitopes required for both monoclonal antibody binding and for T-cell recognition. The data suggest that there are multiple sites on a single Ia molecule that are recognized by T helper cells and also that the tertiary conformation of the Ia molecule can be critical in the formation of such sites.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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