A fluorescence spectroscopic study on the binding of mRNA 5 '-cap-analogs to human translation initiation factor eIF4E: a critical evaluation of the sources of error



: Wieczorek Z, Darzynkiewicz E, Lonnberg H

Publisher: ELSEVIER SCIENCE SA

: 1998

Journal of Photochemistry and Photobiology B: Biology

JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY B-BIOLOGY

: J PHOTOCH PHOTOBIO B

: 43

: 2

: 158

: 163

: 6

: 1011-1344

DOI: https://doi.org/10.1016/S1011-1344(98)00100-6


Equilibrium constants for the association of human protein translation initiation factor eIF4E with two mRNA 5'-cap analogs, namely 7-methylguanosine 5'-triphosphate and P-1-(7-methylguanosine-5') P-3-(guanosine-5') triphosphate, and with guanosine 5'-monophosphate have been redetermined by the fluorescence quenching method taking the inner filter effect of the cap-analog into account. It has been shown that neglecting the latter correction may lead to either underestimation or overestimation of the association constant obtained by applying the Eadie-Hofstee plot: the reasonably firm binding of 7-methylated cap-analogs becomes underestimated, while the weak binding of nonmethylated nucleotides becomes overestimated. (C) 1998 Elsevier Science S.A. All rights reserved.



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