Major and minor epitopes on the self antigen mouse cytochrome c mapped by site-directed mutagenesis

Molecular Immunology
J M MinnerathR Jemmerson

Abstract

Variants of rat (mouse) cytochrome c, prepared by site-directed mutagenesis or represented by closely-related cytochromes c from different species, were employed to map the functional boundaries of a number of mouse monoclonal antibodies (mAb) specific for the major antigenic region on the self antigen (Ag) around residue 62 and the minor antigenic region around residue 44. The recombinant mouse cytochromes c tested were, unlike the tissue-derived Ag, trimethylated at position 72, and included the wild-type which was acetylated at the amino terminus, a variant that was unacetylated at the amino terminus, and variants with the following single amino acid residue replacements: V11I (valine to isoleucine at position 11), Q12M, A15S, A44P, F46Y, D50A, T58I and G89E. Of these, only the A44P variant affected the binding of mAb to the region previously localized to the vicinity of residue 44, thus confirming that assignment. Loss of the acetyl group at the amino terminus affected the binding of most of the mAb to the region around residue 62. The other mutations had little, if any, affect on mAb binding. The epitopes of mAb binding the region around residue 62 were shown in this study to have similar functional boundaries. This site o...Continue Reading

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Citations

May 16, 2006·Apoptosis : an International Journal on Programmed Cell Death·Chad CummingsRonald Jemmerson
Oct 24, 2009·Apoptosis : an International Journal on Programmed Cell Death·Ramil CodinaRonald Jemmerson
Dec 19, 1995·Proceedings of the National Academy of Sciences of the United States of America·J M MinnerathR Jemmerson
Mar 30, 2000·Protein Science : a Publication of the Protein Society·C S RamanB T Nall

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