The antibody-enzyme analogy. Characterization of antibodies to phosphopyridoxyltyrosine derivatives

Biochemistry
V Raso, B D Stollar

Abstract

Stable analogs of the crucial Schiff base intermediate of enzymatic and nonenzymatic pyridoxal phosphate catalysis have been used as haptens for induction of specific antibodies. N-(5-phosphopyridoxyl)-3'-amino-L-tyrosine and its conformationally distinct cyclized derivative resemble the Schiff base formed upon mixing tyrosine with pyridoxal phosphate. These compounds were covalently coupled to a protein carrier via the 3'-amino group so as to confer a prescribed orientation, with the coenzyme region farthest removed from the carrier. A third antigen, with the phosphopyridoxyl group alone as the hapten, was prepared by linkage of pyridoxal phosphate directly to free amino groups on the carrier protein. Antibodies elicited for each determinant were purified by means of appropriate affinity columns. Antibody heterogeneity was observed in that different species could be separated from a given serum by sequential elution from the affinity columns with 1 M sodium phosphate buffers of pH 7.6, 5.2, 2.6 and 1.5. In assays of quantitative precipitation, inhibition of precipitation, equilibrium dialysis, and fluorescence quenching, antibodies to the phosphopyridoxyltyrosine haptens showed specificity for the phosphorylated form of the co...Continue Reading

References

Apr 8, 1985·Journal of Immunological Methods·D L BrandonL L Layton
Jul 1, 1994·Bioorganic & Medicinal Chemistry·D B StephensB L Iverson
Oct 16, 2002·Journal of Immunological Methods·S I GramatikovaP Christen
Oct 16, 2002·Journal of Immunological Methods·Elizabeth L OstlerGerard Gallacher
Jan 1, 1980·Journal of Immunological Methods·J W Thanassi, J A Cidlowski
Dec 30, 2004·Angewandte Chemie·Sergio MartíJuan Bertrán
Oct 1, 1988·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·R A Lerner, S J Benkovic
Oct 17, 2008·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Fang Wu, H Gehring

Related Concepts

Phosphate buffers
Derivatives
Structure-Activity Relationship
Binding Sites, Antibody
Analog
Pyridoxal Phosphate
Plasma Protein Binding Capacity
Staphylococcal Protein A
Chinchilla Rabbits
Antigens

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