Detection of the homology among proteins by immunochemical cross-reactivity between denatured antigens. Application to the threonine and methionine regulated aspartokinases-homoserine dehydrogenases from Escherichia coli K 12.
The two isofunctional enzymes aspartokinases-homoserine dehydrogenases I and II from Escherichia coli K 12 are compared using immunochemical techniques. The antibodies raised against one of these two proteins when in its native state can only recognize the homologous antigen, whether it is native or denatured. Contrarily, the antibodies raised against one of these two proteins when in its denatured state can recognize both the homologous and heterologous denatured antigens. The existence of this cross-reaction only between the two denatured aspartokinases-homoserine dehydrogenases suggests that these two enzymes have some similarity since such a reaction is not detected with several other denatured proteins. The regions involved in this similarity are buried inside the native proteins, and become exposed only upon denaturation. The same results, the existence of a cross-reaction between denatured species and none between the native ones, is obtained with proteolytic fragments derived from these two proteins and endowed with homoserine dehydrogenase activity. This resemblance between the two aspartokinases-homoserine dehydrogenases suggests that these proteins derive from a common ancestor. It is also proposed that such a cross-reaction between two denatured proteins is evidence for an homology between their amino acid sequences, and that the use of denatured proteins as both immunogens and antigens could be useful in detecting sequence homologies.
D000478:Alkylation; D001225:Aspartokinase Homoserine Dehydrogenase / Q000378:metabolism; D003168:Complement Fixation Tests; D003429:Cross Reactions; D004926:Escherichia coli / Q000201:enzymology*; D005033:Ethylmaleimide; D006386:Hemagglutination Tests; D007106:Immune Sera; D007461:Iodoacetates; D007527:Isoenzymes / Q000378:metabolism; D008715:Methionine / Q000494:pharmacology*; D008745:Methylation; D009097:Multienzyme Complexes / Q000378:metabolism; D013912:Threonine / Q000494:pharmacology*
D007106:Immune Sera; D007461:Iodoacetates; D007527:Isoenzymes; D009097:Multienzyme Complexes; D013912:Threonine; D008715:Methionine; D001225:Aspartokinase Homoserine Dehydrogenase; D005033:Ethylmaleimide