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Properties of tRNA-Specific Antibodies from NZB/NZW Mice

From the Laboratory of Chemical Biology and Arthritis and Rheumatism Branch, National Institute of Arthritis, Metabolism, and Digestive Diseases, National Institutes of Health, Bethesda, Maryland 20014

Abstract

Antibodies that bind tRNA are produced spontaneously in New Zealand Black/New Zealand White (NZB/NZW) F₁ hybrid female mice. An assay for the detection of these antibodies has been developed by using gel filtration and radioactive tRNA. This assay was found superior to the widely used ammonium sulfate precipitation assay because of the nature of the interaction between the protein and the tRNA. The antibodies bound native tRNA preferentially to tRNA denatured by cross-linking with formaldehyde. This conformational specificity was confirmed in competition experiments. The antibodies to native tRNA had an average association constant of 5 x 10⁷ liter/mole at 4°C and could bind to more than one site per tRNA molecule. Experiments with immunoglobulin class-specific anti-mouse antisera, in solution and by radioimmunoelectrophoresis, showed that the antibodies were heterogeneous, but were predominantly of the IgG class.

These antibodies may be useful for detection, localization, and conformational analysis of tRNA in solution as well as for understanding the pathogenesis of the lupus-like syndrome in these mice.

Footnotes

¹ Laboratory of Chemical Biology.

² Recipient of a fellowship from Consiglio Nazionale delle Ricerche, Rome, Italy, while on leave from 2nd Chair of Biochemistry, 2nd Medical School, University of Naples, Naples, Italy.

³ Arthritis and Rheumatism Branch.