TY - JOUR
T1 - The purification, crystallisation and preliminary structural characterisation of glucose-1-phosphate thymidylyltransferase (RmlA), the first enzyme of the dTDP-L-rhamnose synthesis pathway from Pseudomonas aeruginosa
AU - Blankenfeldt, W
AU - Giraud, MF
AU - Leonard, G
AU - Rahim, R
AU - Creuzenet, C
AU - Lam, J
AU - Naismith, James Henderson
PY - 2000/11
Y1 - 2000/11
N2 - Glucose-1-phosphate thymidylyltransferase (RmlA; E.C. 2.7.7.24) is the first of four enzymes involved in the biosynthesis of dTDP-L-rhamnose, the precursor of L-rhamnose, a key component of the cell wall of many pathogenic bacteria. RmlA catalyses the condensation of thymidine triphosphate (dTTP) and alpha -D-glucose-1-phosphate (G1P), yielding dTDP-D-glucose. RmlA from Pseudomonas aeruginosa has been overexpressed and purified. Crystals of the enzyme have been grown using the sitting-drop vapour-diffusion technique with PEG 6000 and lithium sulfate as precipitant. Several diffraction data sets of single frozen crystals were collected to a resolution of 1.66 Angstrom. Crystals belonged to space group P1, with unit-cell parameters a = 71.5, b = 73.1, c = 134.7 Angstrom, alpha = 89.9, beta = 80.9, gamma = 81.1 degrees. The asymmetric unit contains eight monomers in the form of two RmlA tetramers with a solvent content of 51%. Selenomethionine-labelled protein has been obtained and crystallized.
AB - Glucose-1-phosphate thymidylyltransferase (RmlA; E.C. 2.7.7.24) is the first of four enzymes involved in the biosynthesis of dTDP-L-rhamnose, the precursor of L-rhamnose, a key component of the cell wall of many pathogenic bacteria. RmlA catalyses the condensation of thymidine triphosphate (dTTP) and alpha -D-glucose-1-phosphate (G1P), yielding dTDP-D-glucose. RmlA from Pseudomonas aeruginosa has been overexpressed and purified. Crystals of the enzyme have been grown using the sitting-drop vapour-diffusion technique with PEG 6000 and lithium sulfate as precipitant. Several diffraction data sets of single frozen crystals were collected to a resolution of 1.66 Angstrom. Crystals belonged to space group P1, with unit-cell parameters a = 71.5, b = 73.1, c = 134.7 Angstrom, alpha = 89.9, beta = 80.9, gamma = 81.1 degrees. The asymmetric unit contains eight monomers in the form of two RmlA tetramers with a solvent content of 51%. Selenomethionine-labelled protein has been obtained and crystallized.
KW - ENTERICA SEROVAR TYPHIMURIUM
KW - 3,5-EPIMERASE
KW - BIOSYNTHESIS
KW - REDUCTASE
KW - PROTEINS
KW - BINDING
UR - http://www.scopus.com/inward/record.url?scp=0033749616&partnerID=8YFLogxK
M3 - Article
SN - 0907-4449
VL - D56
SP - 1501
EP - 1504
JO - Acta Crystallographica. Section D, Biological crystallography
JF - Acta Crystallographica. Section D, Biological crystallography
ER -