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

Acta Crystallogr D Biol Crystallogr. 2000 Sep;56(Pt 9):1176-9. doi: 10.1107/s0907444900008519.

Abstract

Carbonic anhydrases are zinc metalloenzymes that fall into three distinct evolutionary and structural classes, alpha, beta and gamma. Although alpha-class enzymes, particularly mammalian carbonic anhydrase II, have been the subject of extensive structural studies, for the beta class, consisting of a wide variety of prokaryotic and plant chloroplast carbonic anhydrases, the structural data is quite limited. A member of the beta 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.

MeSH terms

  • 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

Substances

  • Bacterial Proteins
  • Cations
  • Isoenzymes
  • Selenomethionine
  • Carbonic Anhydrases
  • Selenium
  • Zinc