Crystallization of diaminopimelate decarboxylase from Escherichia coli, a stereospecific D-amino-acid decarboxylase.
The final step in lysine biosynthesis in bacteria, the conversion of meso-diaminopimelate to L-lysine, is catalyzed by the only known D-amino-acid decarboxylase, diaminopimelate decarboxylase (DDC). The Escherichia coli DDC has been cloned, overexpressed in E. coli with a carboxy-terminal polyhistidine purification tag and crystallized from lithium sulfate. The protein is intensely yellow, owing to the pyridoxal-5'-phosphate cofactor, and is enzymatically active. Large well ordered crystals, belonging to space group P6(1)22 with unit-cell parameters a = b = 98.6, c = 177 A, make high-resolution X-ray diffraction studies possible to characterize the residues important in stereospecific decarboxylation and reprotonation during catalytic turnover.
Momany
Cory
C
Department of Pharmaceutical and Biomedical Sciences, Wilson College of Pharmacy, University of Georgia, Athens, Georgia, USA. cmomany@mail.rx.uga.edu
Levdikov
Vladimir
V
Blagova
Lena
L
Crews
Kristine
K
D001426:Bacterial Proteins; D002262:Carboxy-Lyases / Q000737:chemistry* / Q000235:genetics; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D003653:Decarboxylation; D004926:Escherichia coli / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D011487:Protein Conformation
D001426:Bacterial Proteins; D002262:Carboxy-Lyases; C079912:LysA protein, Bacteria; C020031:diaminopimelic acid decarboxylase