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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

✍ Scribed by Zbigniew Wieczorek; Edward Darzynkiewicz; Harri Lönnberg


Publisher
Elsevier Science
Year
1998
Tongue
English
Weight
549 KB
Volume
43
Category
Article
ISSN
1011-1344

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✦ Synopsis


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'-(7-methylguanosine-5') P'-(guanosine-5') triphosphate, and with guanosine 5'-monophosphate have been redetermined by the fluorescence quenching method taking the inner filter effect of the cup-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 cup-analogs becomes underestimated, while the weak binding of nonmethylated nucleotides becomes overestimated.