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Table 1 Kinetic parameters of MATs from different domains of life

From: Structural and functional characterisation of the methionine adenosyltransferase from Thermococcus kodakarensis

 

Tk MAT

Mj MAT

Ec MAT

Hs MAT

kinetic parameters

    

V max [μmol/min/mg]

1.95 ± 0.26

3.0

1.2

0.2

K M (Met) [mM]

0.31 ± 0.07

0.14

0.08

0.003

K M (ATP) [mM]

6.54 ± 2.49

0.25

0.11

0.03

assay

HPLC (SCX column)

radioactive filter binding assay

radioactive filter binding assay

radioactive filter binding assay

buffer

0.1 M Tris–HCl, 0.02 M MgCl2, 0.2 KCl, pH 8.0

50 mM K-Hepes, 10 mM MgCl2, 25 mM KCl, pH 8.0

0.1 Tris-Cl, 6 mM MgCl2, 0.1 KCl, pH 8.3

50 mM K-Tes, 50 mM KCl, 10 mM MgCl2, 10 mM DTT, 0.3 mM Na2EDTA, 0.1 mM BSA, pH 7.4

substrates

10 mM l-methionine 10 mM ATP

0.1 mM l-[methyl-14C]-methionine 5 mM ATP

0.6 mM l-[methyl-14C]-methionine 4.4 mM 14C-ATP

0.01 mM l-[methyl-14C]-methionine 5 mM ATP

temperature

37°C

70°C

25°C

37°C

reference

this work

Graham et al. [16]

Markham et al. [12]

Kotb & Kredich [25]