Cloning, crystallization and preliminary characterization of a β-carbonic anhydrase from Escherichia coli

• Published in: Acta crystallographica. Section D, Biological crystallography. Biological crystallography., 2000-09, Vol.56 (9), p.1176-1179
• Main Authors: Cronk, Jeff D., O'Neill, Jason W., Cronk, Michelle R., Endrizzi, James A., Zhang, Kam Y. J.
• Format: Article
• Language: English
• Subjects: Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Carbonic Anhydrases - chemistry; Carbonic Anhydrases - genetics; Cations; Cloning, Molecular; Crystallization; Crystallography, X-Ray; Escherichia coli - enzymology; Escherichia coli - genetics; Isoenzymes - chemistry; Isoenzymes - genetics; Selenium - chemistry; Selenomethionine - chemistry; Zinc - chemistry; zinc metalloenzymes; β-carbonic anhydrases
• ISSN: 1399-0047; 0907-4449; 1399-0047
• DOI: 10.1107/S0907444900008519

Carbonic anhydrases are zinc metalloenzymes that fall into three distinct evolutionary and structural classes, α, β and γ. Although α‐­class enzymes, particularly mammalian carbonic anhydrase II, have been the subject of extensive structural studies, for the β class, consisting of a wide variety of prokaryotic and plant chloroplast carbonic anhydrases, the structural data is quite limited. A member of the β class from E. coli (CynT2) has been crystallized in native and selenomethionine‐labelled forms and multiwavelength anomalous dispersion techniques have been applied in order to determine the positions of anomalous scatterers. The resulting phase information is sufficient to produce an interpretable electron‐density map. A crystal structure for CynT2 would contribute significantly to the emerging structural knowledge of a biologically important class of enzymes that perform critical functions in carbon fixation and prokaryotic metabolism.