title: Crystallization of Mycobacterium smegmatis methionyl-tRNA synthetase in the presence of methionine and adenosine.
abstract: Methionyl-tRNA synthetase (MetRS) from Mycobacterium smegmatis was recombinantly expressed in Escherichia coli and purified using Ni(2+)-affinity and size-exclusion chromatography. Crystals formed readily in the presence of the ligands methionine and adenosine. These two ligands are components of an intermediate in the two-step catalytic mechanism of MetRS. The crystals were produced using the vapour-diffusion method and a full data set to 2.1 A resolution was collected from a single crystal. The crystal belonged to the monoclinic space group C2, with unit-cell parameters a = 155.9, b = 138.9, c = 123.3 A, beta = 124.8 degrees . The presence of three molecules in the asymmetric unit corresponded to a solvent content of 60% and a Matthews coefficient of 3.1 A(3) Da(-1). Structure determination is in progress.
authors.affiliation: Department of Cell and Molecular Biology, Uppsala University, Biomedical Center, Box 596, SE-751 24 Uppsala, Sweden.
authors.forename: Henrik
authors.initials: H
authors.lastname: Ingvarsson
authors.forename: T Alwyn
authors.initials: TA
authors.lastname: Jones
authors.forename: Torsten
authors.initials: T
authors.lastname: Unge
mesh_terms: D000241:Adenosine / Q000737:chemistry*; D000595:Amino Acid Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D003460:Crystallization; D003625:Data Collection; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D008024:Ligands; D008715:Methionine / Q000737:chemistry*; D008718:Methionine-tRNA Ligase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D020102:Mycobacterium smegmatis / Q000201:enzymology*; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013223:Statistics as Topic; D014169:Transformation, Bacterial; D014961:X-Ray Diffraction
chemical_list: D008024:Ligands; D011994:Recombinant Proteins; D008715:Methionine; D008718:Methionine-tRNA Ligase; D000241:Adenosine