Cloning, crystallization and preliminary characterization of a beta-carbonic anhydrase from Escherichia coli.
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.
Cronk
J D
JD
Division of Basic Sciences, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, Seattle, WA 98109-1024, USA.
O'Neill
J W
JW
Cronk
M R
MR
Endrizzi
J A
JA
Zhang
K Y
KY
D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D002256:Carbonic Anhydrases / Q000737:chemistry* / Q000235:genetics; D002412:Cations; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D007527:Isoenzymes / Q000737:chemistry* / Q000235:genetics; D012643:Selenium / Q000737:chemistry; D012645:Selenomethionine / Q000737:chemistry; D015032:Zinc / Q000737:chemistry
D001426:Bacterial Proteins; D002412:Cations; D007527:Isoenzymes; D012645:Selenomethionine; D002256:Carbonic Anhydrases; D012643:Selenium; D015032:Zinc