Purification, crystallization and preliminary crystallographic studies of plant S-adenosyl-L-homocysteine hydrolase (Lupinus luteus).

By degrading S-adenosyl-L-homocysteine, which is a byproduct of S-adenosyl-L-methionine-dependent methylation reactions, S-adenosyl-L-homocysteine hydrolase (SAHase) acts as a regulator of cellular methylation processes. S-Adenosyl-L-homocysteine hydrolase from the leguminose plant yellow lupin (Lupinus luteus), LlSAHase, which is composed of 485 amino acids and has a molecular weight of 55 kDa, has been cloned, expressed in Escherichia coli and purified. Crystals of LlSAHase in complex with adenosine were obtained by the hanging-drop vapour-diffusion method using 20%(w/v) PEG 4000 and 10%(v/v) 2-propanol as precipitants in 0.1 M Tris-HCl buffer pH 8.0. The crystals were tetragonal, space group P4(3)2(1)2, with unit-cell parameters a = 122.4, c = 126.5 A and contained two protein molecules in the asymmetric unit, corresponding to the functional dimeric form of the enzyme. Atomic resolution (1.17 A) X-ray diffraction data have been collected using synchrotron radiation.