D214A mutant of tyrosine phenol-lyase from Citrobacter freundii

Crystal Structure of Citrobacter Freundii Asp214Ala Tyrosine Phenol-Lyase Reveals that Asp214 is Critical for Maintaining a Strain in the Internal Aldimine

SINCE THE UNIT-CELL PARAMETERS FOR D214A TPL CRYSTAL WERE ONLY SLIGHTLY DIFFERENT THAN FOR THE NATIVE HOLOENZYME, IT WAS POSSIBLE TO REFINE THE D214A TPL MUTANT STRUCTURE IN REFMAC USING THE WILD-TYPE TPL HOLOENZYME STRUCTURE, PDB ENTRY 2EZ1, AS A STARTING MODEL.

THE CRYSTALS WERE GROWN AT 293 K BY THE HANGING DROP VAPOR DIFFUSION TECHNIQUE BY MIXING 2 UL OF THE PROTEIN SOLUTION, 18-20 MG/ML, WITH AN EQUAL VOLUME OF THE RESERVOIR SOLUTION CONTAINING 50 MMOL/L TRIETHANOLAMINE BUFFER PH 8.0, 0.4-0.8 MOL/L KCL, 0.35-0.38 G/ML PEG 5000 MME, 5 MMOL/L PYRIDOXAL-5-PHOSPHATE, AND 2 MMOL/L DITHIOTHREITOL.

HOLO TYROSINE PHENOL-LYASE FROM CITROBACTER FREUNDII AT PH8.0

APO TYROSINE PHENOL-LYASE FROM CITROBACTER FREUNDII AT PH8.0

TYROSINE PHENOL-LYASE FROM CITROBACTER INTERMEDIUS COMPLEX WITH 3-(4'-HYDROXYPHENYL)PROPIONIC ACID, PYRIDOXAL-5 '-PHOSPHATE AND CS+ ION

QUINONOID INTERMEDIATE OF CITROBACTER FREUNDII TYROSINE PHENOL-LYASE FORMED WITH ALANINE

QUINONOID INTERMEDIATE OF CITROBACTER FREUNDII TYROSINE PHENOL-LYASE FORMED WITH METHIONINE

Y71F MUTANT OF TYROSINE PHENOL-LYASE FROM CITROBACTER FREUNDII IN COMPLEX WITH QUINONOID INTERMEDIATE FORMED WITH 3-FLUORO-L-TYROSINE

F448H MUTANT OF TYROSINE PHENOL-LYASE FROM CITROBACTER FREUNDII IN COMPLEX WITH QUINONOID INTERMEDIATE FORMED WITH 3-FLUORO-L-TYROSINE

TYROSINE PHENOL-LYASE FROM CITROBACTER FREUNDII IN COMPLEX WITH PYRIDINE N-OXIDE AND THE QUINONOID INTERMEDIATE FORMED WITH L-ALANINE

ENGINEERED RESIDUE IN CHAIN A, ASP 214 TO ALA ENGINEERED RESIDUE IN CHAIN B, ASP 214 TO ALA

HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS.

LYASE, PYRIDOXAL-5'-PHOSPHATE-DEPENDENT ENZYME, BETA-ELIMINATION REACTION