论文信息 - Crystallization and preliminary X-ray analysis of the Escherichia coli UDP-MurNAc-tripeptide D-alanyl-D-alanine-adding enzyme (MurF).

Crystallization and preliminary X-ray analysis of the Escherichia coli UDP-MurNAc-tripeptide D-alanyl-D-alanine-adding enzyme (MurF).

Crystals of the Escherichia coli UDP-MurNAc-tripeptide D-Ala-D-Ala-adding protein (MurF), which catalyzes the formation of the last metabolite of the bacterial cell-wall building block, have been grown in hanging-drop vapor-diffusion trials using PEG 8K as a precipitating agent. The crystals belong to hexagonal space group P6(1) or P6(5), with unit-cell dimensions a = b = 74, c = 425 A. The asymmetric unit contains two molecules, with a crystal volume per protein mass (V(m)) of 3.4 A(3) Da(-1) and a solvent content of about 64% by volume. A native data set to 2.8 A resolution has been obtained from a frozen crystal using a synchrotron X-ray source.

[1] W Furey,et al. PHASES-95: a program package for processing and analyzing diffraction data from macromolecules. , 1997, Methods in enzymology.

[2] B. Leiting,et al. High-level expression of soluble protein in Escherichia coli using a His6-tag and maltose-binding-protein double-affinity fusion system. , 1997, Protein expression and purification.

[3] E. Fanchon,et al. Crystal structure of UDP‐N‐acetylmuramoyl‐L‐alanine:D‐glutamate ligase from Escherichia coli , 1997, The EMBO journal.

[4] J. Sambrook,et al. Molecular Cloning: A Laboratory Manual , 2001 .

[5] Z. Otwinowski,et al. [20] Processing of X-ray diffraction data collected in oscillation mode. , 1997, Methods in enzymology.

[6] D. Pompliano,et al. Kinetic mechanism of the Escherichia coli UDPMurNAc-tripeptide D-alanyl-D-alanine-adding enzyme: use of a glutathione S-transferase fusion. , 1996, Biochemistry.

[7] C. Walsh,et al. Vancomycin resistance: structure of D-alanine:D-alanine ligase at 2.3 A resolution. , 1994, Science.

[8] B. Matthews. Solvent content of protein crystals. , 1968, Journal of molecular biology.