Crystal structure of K. lactis Edc1-Dcp1-Dcp2-Edc3 decapping complex with synthetic cap substrate analog

[[(2~{R},3~{S},4~{R},5~{R})-5-(2-azanyl-7-methyl-6-oxidanylidene-3~{H}-purin-7-ium-9-yl)-3,4-bis(oxidanyl)oxolan-2-yl]methoxy-sulfanyl-phosphoryl] [[[(2~{R},3~{S},4~{R},5~{R})-5-(2-azanyl-7-methyl-6-oxidanylidene-3~{H}-purin-7-ium-9-yl)-3,4-bis(oxidanyl)oxolan-2-yl]methoxy-sulfanyl-phosphoryl]oxy-oxidanyl-phosphoryl] hydrogen phosphate

Structure of the activated Edc1-Dcp1-Dcp2-Edc3 mRNA decapping complex with substrate analog poised for catalysis.

The distance between Residue F PHE 128 and Residue F ASN 131 is 14.50 Angstrom. But there are only 2 residues (not enough sequence) to cover the gap region.

Water-cooled flat double Si(111) Khozu monochromator

Kluyveromyces lactis (strain ATCC 8585 / CBS 2359 / DSM 70799 / NBRC 1267 / NRRL Y-1140 / WM37)

Protein solution: 4mg/mL Kl Dcp1/Dcp2/Edc3, 1mM Edc1 peptide, 6mM substrate analog, 10mM magnsium chloride.
Well solution: 0.22M magnesium chloride, 0.03M EDTA, 8% PEG 8000.
Procedure: 250nL protein solution + 250nL well solution + 50nL 1:10000 seed stock. Set up at RT, grow crystals at 4C.

National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)

mRNA decay, decapping, Nudix, nucleotide analog, TRANSLATION