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Determination of the base-pairing content of four specific tRNAs by infrared spectroscopy

✍ Scribed by U. Schernau; Th. Ackermann

Publisher
Wiley (John Wiley & Sons)
Year
1977
Tongue
English
Weight
448 KB
Volume
16
Category
Article
ISSN
0006-3525

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✦ Synopsis

Abstract

Infrared spectroscopy was used for the determination of the base‐pairing content of four specific tRNAs in deuterium oxide solution. Infrared spectra were obtained in the 1750–1550 cm^−1^ region at various temperatures ranging from about 15 to 90°C. Melting curves were constructed by plotting the molar extinction coefficient at ν = 1657 cm^−1^ versus temperature. These transition curves enabled us to determine the ranges of temperature which correspond to the ordered (partially double‐stranded) or randomly coiled structure of the tRNA. For a set of wavenumbers the extinction coefficients at these temperatures were used for the calculation of the base‐pairing content. The procedure employed here is based on a method described earlier by Thomas [(1969) Biopolymers 7, 325–334]. For the conditions selected for this investigation (Mg^2+^‐free D~2~O‐buffer; 0.01__M__ tris‐DCl, 0.015__M__ NaCl, pD 7.5) the results of this determination agree within the limits of errors with the number of base pairs predicted by the cloverleaf model.

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