X-ray structure of beta-carbonic anhydrase from the red alga, Porphyridiumpurpureum, reveals a novel catalytic site for CO2 hydration

The carbonic anhydrases (CAs) fall into three evolutionarily distinct famil ies designated alpha-, beta-, and gamma-CAs based on their primacy structur e. beta-CAs are present in higher plants, algae, and prokaryotes, and are i nvolved in inorganic carbon utilization, Here, we describe the novel x-ray structure of beta-CA from the red alga, Porphyridium purpureum, at 2.2-Angs trom resolution using intrinsic zinc multiwavelength anomalous diffraction phasing. The CA monomer is composed of two internally repeating structures, being folded as a pair of fundamentally equivalent motifs of an alpha/beta domain and three projecting alpha-helices. The motif is obviously distinct from that of either alpha- or gamma-CAs. This homodimeric CA appears like a tetramer with a pseudo 222 symmetry. The active site zinc is coordinated by a Cys-Asp-His-Cys tetrad that is strictly conserved among the beta-CAs, No water molecule is found in a zinc-liganding radius, indicating that the zinc-hydroxide mechanism in alpha-CAs, and possibly in gamma-CAs, is not di rectly applicable to the case in beta-CAs. Zinc coordination environments o f the CAs provide an interesting example of the convergent evolution of dis tinct catalytic sites required for the same CO2 hydration reaction.