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Formation of an Antibody with Serologic Specificity for N⁶-(²-Isopentenyl) Adenosine¹

From the Department of Pharmacology and Toxicology, University of Rochester Medical Center, Rochester, New York 14642 and the Graduate Department of Biochemistry, Brandeis University, Waltham, Massachusetts 02154

Abstract

Recent reports have dealt with the demonstration of specific antibodies found in response to immunization of rabbits with conjugates of methylated nucleoside haptens (1, 2). Not surprising were the findings that the methyl group and its position on the purine or pyrimidine ring dominated the degree of complementarity between hapten and antibody. Nevertheless, some recognition by these antibodies of heterologous methylated bases and even normal bases was observed. We have recently produced in rabbits antibodies directed toward N⁶-(²-isopentenyl) adenosine (iA). As expected, the N⁶-(²-isopentenyl) group on the nucleoside was immunodominant. These antibodies do react with N⁶-methyladenosine; yet 1 x 10⁴ more N⁶-methyladenosine molecules were required for equivalent activity. Even at a 1 x 10⁵ higher concentration, normal bases did not cross-react. The iA in the tRNA was available for reaction with the antibody of iA.

Isolation and characterization of the iA used for synthesis of the immunizing antigens have been described previously (3).

Footnotes

¹ This investigation was supported in part by grants from the National Institutes of Health (Cancer Center Grant CA-11198) and the American Cancer Society (Grant IN-18K) and by Public Health Service Research Grant CA-12269 to Bruce Hacker and Contract 71-2163 from the National Cancer Institute to Lawrence Levine. This work is Publication 819 from the Graduate Department of Biochemistry, Brandeis University.

² American Cancer Society Professor of Biochemistry (Award PRP-21).

³ Public Health Service Research Career Awardee (Award 5K6A12372).