NMR Structure of a tRNA 2'-phosphotransferase from Runella slithyformis in complex with NAD+

tRNA 2'-phosphotransferase (E.C.2.7.1.160)

[(2R,3S,4R,5R)-5-(3-aminocarbonylpyridin-1-ium-1-yl)-3,4-dihydroxy-oxolan-2-yl]methyl [[(2R,3S,4R,5R)-5-(6-aminopurin-9-yl)-3,4-dihydroxy-oxolan-2-yl]methoxy-hydroxy-phosphoryl] phosphate

NMR solution structures of Runella slithyformis RNA 2'-phosphotransferase Tpt1 provide insights into NAD+ binding and specificity.

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600 uM [U-13C; U-15N] Tpt1, 2.4 mM NICOTINAMIDE-ADENINE-DINUCLEOTIDE,
20 mM HEPES, 200 mM sodium chloride, 2 mM DTT, 2 mM [U-2H] EDTA,
5 % v/v [U-2H] glycerol, 1 mM AEBSF protease inhibitor,
0.04 % w/v sodium azide, 50 uM DSS, 95% H2O/5% D2O

280 uM [U-13C; U-15N] Tpt1, 1.12 mM NICOTINAMIDE-ADENINE-DINUCLEOTIDE,
20 mM HEPES, 200 mM sodium chloride, 2 mM DTT, 2 mM [U-2H] EDTA,
5 % v/v [U-2H] glycerol, 1 mM AEBSF protease inhibitor,
0.04 % w/v sodium azide, 50 uM DSS, 95% H2O/5% D2O

325 uM [U-13C; U-15N] Tpt1, 1.3 mM NICOTINAMIDE-ADENINE-DINUCLEOTIDE,
20 mM HEPES, 200 mM sodium chloride, 2 mM DTT, 2 mM [U-2H] EDTA,
5 % v/v [U-2H] glycerol, 1 mM AEBSF protease inhibitor,
0.04 % w/v sodium azide, 50 uM DSS, 100% D2O

300 uM [U-13C; U-15N] Tpt1, 1.2 mM NICOTINAMIDE-ADENINE-DINUCLEOTIDE,
20 mM HEPES, 200 mM sodium chloride, 2 mM DTT, 2 mM [U-2H] EDTA,
5 % v/v [U-2H] glycerol, 1 mM AEBSF protease inhibitor,
0.04 % w/v sodium azide, 50 uM DSS, 100% D2O

500 uM [U-13C; U-15N; U-2H] Tpt1, 2 mM NICOTINAMIDE-ADENINE-DINUCLEOTIDE,
20 mM HEPES, 200 mM sodium chloride, 2 mM DTT, 2 mM [U-2H] EDTA,
5 % v/v [U-2H] glycerol, 1 mM AEBSF protease inhibitor,
0.04 % w/v sodium azide, 50 uM DSS, 95% H2O/5% D2O

400 uM [U-13C; U-15N] Tpt1, 1.6 mM NICOTINAMIDE-ADENINE-DINUCLEOTIDE,
20 mM HEPES, 200 mM sodium chloride, 2 mM DTT, 2 mM [U-2H] EDTA,
5 % v/v [U-2H] glycerol, 1 mM AEBSF protease inhibitor,
0.04 % w/v sodium azide, 50 uM DSS, 100% D2O

Schwieters, Kuszewski, Tjandra and Clore

Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax