5’-methylthioadenosine/S-adenosylhomocysteine nucleosidase (MTAN) plays a critical role in diverse pathways in bacterial cells such as biological methylation, polyamine biosynthesis, methionine recycling, and bacterial quorum sensing. It has been known that MtaN catalyzes the hydrolysis of N-ribosidic bond of adenosine-based substrates such as S-adenosyl-L-homocysteine (SAH), S-methyl-5’-thioadenosine (MTA) and 5’-deoxyadenosine (5’-DOA). In Aeromonas hydrophila, there are two MtnN subfamily proteins: MtaN-1, a periplasmic protein with an N-terminal signal peptide; and MtaN-2, a cytosolic protein. In this study, MtaN-2 from A. hydrophila was successfully expressed and purified using Ni-NTA affinity, Q anion-exchange, and gel-filtration chromatography. We first crystallized apo MtaN-2 but it diffracted to a low resolution of 5.1 Å. New crystals suitable for diffraction were obtained by adding 2 mM adenosine, a substrate analog of MtaN-2 during purification process and the crystals diffracted to the resolution of 2.0 Å. The crystals belong to the trigonal space group P31 or P32, with unit-cell parameters of a = b = 74.94 Å and c = 185.21 Å. The asymmetric unit contains four complexes of MtaN-2 with hydrolysis products of adenosine.
INTRODUCTION
RESULTS AND DISCUSSION
METHODS