1. Crystallization of diaminopimelate decarboxylase from Escherichia coli, a stereospecific D-amino-acid decarboxylase
- Author
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Lena Blagova, Vladimir Levdikov, Kristine Crews, and Cory Momany
- Subjects
Decarboxylation ,Carboxy-Lyases ,Protein Conformation ,medicine.disease_cause ,Crystallography, X-Ray ,Diaminopimelate decarboxylase ,Cofactor ,Catalysis ,law.invention ,Stereospecificity ,Bacterial Proteins ,Structural Biology ,law ,medicine ,Escherichia coli ,Crystallization ,Cloning, Molecular ,biology ,General Medicine ,biology.organism_classification ,Biochemistry ,biology.protein ,bacteria ,Bacteria - Abstract
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.
- Published
- 2001