Expression, purification, crystallization and preliminary X-ray analysis of Escherichia coli 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase.
A recombinant form of Escherichia coli 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase (E.C. 3.2.2.9) has been purified to homogeneity and crystallized using the hanging-drop vapour-diffusion technique. While several different crystallization conditions were obtained, only one set of conditions yielded crystals suitable for X-ray diffraction analysis. These crystals grow as diamond-shaped wedges, with unit-cell parameters a = 50.92, b = 133.99, c = 70.88 A, alpha = beta = gamma = 90 degrees. The crystals belong to space group P2(1)2(1)2 and diffract to a minimum d spacing of 2.3 A on a MAR345 image plate with a Rigaku RU-200 rotating-anode X-ray generator. On the basis of density calculations, two monomers are predicted per asymmetric unit (Matthews coefficient, V(M) = 2.37 A(3) Da(-1)), with a solvent content of 48%.
Lee
J E
JE
Structural Biology and Biochemistry, Research Institute, Hospital for Sick Children, 555 University Avenue, Toronto, Ontario M5G 1X8, Canada.
Cornell
K A
KA
Riscoe
M K
MK
Howell
P L
PL
D000595:Amino Acid Sequence; D003460:Crystallization; D018360:Crystallography, X-Ray; D003839:Deoxyadenosines / Q000737:chemistry*; D004926:Escherichia coli / Q000201:enzymology; D008969:Molecular Sequence Data; D009699:N-Glycosyl Hydrolases / Q000737:chemistry*; D013872:Thionucleosides / Q000737:chemistry*
D003839:Deoxyadenosines; D013872:Thionucleosides; C008500:5'-methylthioadenosine; D009699:N-Glycosyl Hydrolases; C019606:adenosylhomocysteine nucleosidase