Structural similarities among enzyme pterin binding sites as demonstrated by a monoclonal anti-idiotypic antibody.
BALB/c mice were immunized with a synthetic co-factor of the aromatic amino acid hydroxylases, 6,7-dimethyl-5,6,7,8-tetrahydropterin, conjugated to albumin. Hybridoma cell lines isolated from the immunized mice secreted monoclonal antibodies reacting specifically with the pterin molecule and monoclonal antibodies which were found to bind phenylalanine hydroxylase. Several lines of evidence were consistent with the anti-phenylalanine hydroxylase antibodies being anti-idiotype antibodies mimicking the pterin molecule and binding to the pterin binding site of phenylalanine hydroxylase. (a) An anti-idiotype monoclonal antibody, NS7, when reimmunized into mice produced anti-pterin antibodies consistent with NS7 being an internal image anti-idiotypic antibody. (b) NS7 antibody was prevented from binding to phenylalanine hydroxylase when a competitive inhibitor of phenylalanine hydroxylase enzyme activity, 6,7-dimethyl-7,8-dihydropterin, was bound to phenylalanine hydroxylase. (c) NS7 antibody was shown to bind to a wide range of pterin-requiring enzymes: phenylalanine, tyrosine and tryptophan hydroxylases, dihydropteridine reductase, dihydrofolate reductase, and sepiapterin reductase. Thus the NS7 antibody has successfully mimicked a common portion of the pterin cofactors utilized by these enzymes and demonstrated structure homology in their pterin binding sites despite their diverse function and little amino acid sequence homology except among the three aromatic amino acid hydroxylases.
Jennings
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Olive Miller Protein Laboratory, Murdoch Institute, Royal Children's Hospital, Melbourne, Victoria, Australia.
Cotton
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R
D000311:Adrenal Glands / Q000201:enzymology; D000429:Alcohol Oxidoreductases / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex / Q000032:analysis; D001665:Binding Sites; D002417:Cattle; D004093:Dihydropteridine Reductase / Q000276:immunology / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007130:Immunoglobulin Idiotypes; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D010651:Phenylalanine Hydroxylase / Q000276:immunology / Q000378:metabolism*; D011621:Pteridines / Q000276:immunology*; D051381:Rats; D013762:Tetrahydrofolate Dehydrogenase / Q000276:immunology / Q000378:metabolism*; D014365:Tryptophan Hydroxylase / Q000276:immunology / Q000378:metabolism*; D014446:Tyrosine 3-Monooxygenase / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D007130:Immunoglobulin Idiotypes; D011621:Pteridines; C005063:2-amino-4-hydroxy-6,7-dimethyl-5,6,7,8-tetrahydropteridine; D000429:Alcohol Oxidoreductases; C022630:sepiapterin reductase; D010651:Phenylalanine Hydroxylase; D014446:Tyrosine 3-Monooxygenase; D014365:Tryptophan Hydroxylase; D013762:Tetrahydrofolate Dehydrogenase; D004093:Dihydropteridine Reductase; D009247:NADH, NADPH Oxidoreductases