Phosphorylation of eIF4E attenuates its interaction with mRNA 5′ cap analogs by electrostatic repulsion: Intein-mediated protein ligation strategy to obtain phosphorylated protein

TABLE 1.

Equilibrium association constants (Kas) for interactions of phosphorylated and unphosphorylated eIF4E with various cap analogs at 20°C

Kas [μM−1]
Cap analogs eIF4E (28–217) eIF4E (28–217) T205C eIF4E (28–217) T205C/S209phos Kas (wt)/Kas (phos)
m7GTP 119.7 ± 5.7 111.6 ± 3.7 42.6 ± 1.8 2.8
m7GDP 20.9 ± 1.5 17.8 ± 0.6 10.5 ± 0.3 2.0
m7GMP 1.5 ± 0.1 1.5 ± 0.1 1.1 ± 0.1 1.4
m7GpppG 12.5 ± 0.3 10.5 ± 0.3 5.8 ± 0.3 2.2
m27,3′OGpppG 10.2 ± 0.3 7.9 ± 0.2 4.33 ± 0.15 2.4
m7Gpppm2′OG 7.5 ± 0.2 7.8 ± 0.2 3.2 ± 0.2 2.3
m7GpppC 6.1 ± 0.3 5.9 ± 0.2 2.8 ± 0.2 2.2
m7GpppA 5.0 ± 0.2 4.9 ± 0.1 2.26 ± 0.10 2.2
m7Gpppm6A 3.9 ± 0.2 4.25 ± 0.20 1.99 ± 0.70 2.0
m7Gpppm7G 5.0 ± 0.2 4.83 ± 0.20 2.84 ± 0.10 1.8
m7GppppG 110.9 ± 6.0 97.5 ± 3.2 27.3 ± 1.1 4.1
m7Gppppm7G 49.4 ± 1.5 50.8 ± 1.6 20.1 ± 1.0 2.5
m7GpppAm2′pUm2′pAm2′ 43.0 ± 1.8 44.0 ± 2.6 9.6 ± 0.4 4.5

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  1. RNA 9: 52-61

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