2.6 Determination of influences of different forms of H protein
Different forms of H proteins (Hapo,
Hlip, HK64A) were used to investigate
their effects on the reactions catalyzed by LplA. The reaction mixture
(200 µL) contained 2 mM lipoic acid, 2 mM ATP, 2 mM
MgCl2, 10 μΜ LplA, Tris-HCl (50 mM, pH 7.0) and
different concentrations of H proteins (from 0.4 to 400 μM). The
components except ATP were premixed and centrifuged before the reactions
were initiated by adding ATP to the reaction mixture.