PDBsum entry 2w3q

Lyase

PDB id: 2w3q

Contents

Protein chain

229 a.a. *

Metals

_CL

_ZN

Waters ×165

* Residue conservation analysis

PDB id:

2w3q

Name:

Lyase

Title:

Structure and inhibition of the co2-sensing carbonic anhydrase can2 from the pathogenic fungus cryptococcus neoformans

Structure:

Carbonic anhydrase 2. Chain: a. Synonym: carbonic anhydrase. Engineered: yes

Source:

Cryptococcus neoformans. Organism_taxid: 5207. Variant: grubii. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

1.34Å R-factor: 0.139 R-free: 0.185

Authors:

C.Schlicker,R.A.Hall,D.Vullo,S.Middelhaufe,M.Gertz,C.T.Supuran, F.A.Muehlschlegel,C.Steegborn

Key ref:

C.Schlicker et al. (2009). Structure and Inhibition of the CO(2)-Sensing Carbonic Anhydrase Can2 from the Pathogenic Fungus Cryptococcus neoformans. J Mol Biol, 385, 1207-1220. PubMed id: 19071134 DOI: 10.1016/j.jmb.2008.11.037

Date:

14-Nov-08 Release date: 30-Dec-08

Protein chain

Q3I4V7  (Q3I4V7_CRYNV) -  Carbonic anhydrase from Cryptococcus neoformans var. grubii

Seq: 239 a.a.

Struc: 229 a.a.

Enzyme reactions

• Enzyme class: E.C.4.2.1.1 - carbonic anhydrase.
• Reaction: hydrogencarbonate + H⁺ = CO2 + H2O
• Cofactor: Zn(2+)

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

Added reference

DOI no: 10.1016/j.jmb.2008.11.037

J Mol Biol 385:1207-1220 (2009)

PubMed id: 19071134

Structure and Inhibition of the CO(2)-Sensing Carbonic Anhydrase Can2 from the Pathogenic Fungus Cryptococcus neoformans.

C.Schlicker, R.A.Hall, D.Vullo, S.Middelhaufe, M.Gertz, C.T.Supuran, F.A.Mühlschlegel, C.Steegborn.

ABSTRACT

In the pathogenic fungus Cryptococcus neoformans, a CO(2)-sensing system is essential for survival in the natural environment ( approximately 0.03% CO(2)) and mediates the switch to virulent growth in the human host ( approximately 5% CO(2)). This system is composed of the carbonic anhydrase (CA) Can2, which catalyzes formation of bicarbonate, and the fungal, bicarbonate-stimulated adenylyl cyclase Cac1. The critical role of these enzymes for fungal metabolism and pathogenesis identifies them as targets for antifungal drugs. Here, we prove functional similarity of Can2 to the CA Nce103 from Candida albicans and describe its biochemical and structural characterization. The crystal structure of Can2 reveals that the enzyme belongs to the "plant-type" beta-CAs but carries a unique N-terminal extension that can interact with the active-site entrance of the dimer. We further tested a panel of compounds, identifying nanomolar Can2 inhibitors, and present the structure of a Can2 complex with the inhibitor and product analog acetate, revealing insights into interactions with physiological ligands and inhibitors.

Selected figure(s)

Figure 3.
Fig. 3. Active site and inhibition of Can2. (a) Active site of Can2 with electron density contoured at 1.0 σ. Coordination of the active-site Zn^2+ by Cys68, His124, Cys127, and a water molecule is indicated by black broken lines, and the hydrogen bond from the water molecule to Asp70 is colored orange. (b) Overlay of the active sites of Can2 and the plant-type β-CAs of E. coli (yellow) and P. sativum (dark red) and the Cab-type β-CAs of M. thermoautotrophicum (green) and M. tuberculosis Rv1284 (gray). The Zn^2+ ions are shown as orange spheres, and the water molecules are colored according to the organism. (c) Chemical structure of the most potent inhibitors identified for Can2, AAZ, and benzolamide. (d) Detrimental effect of the Can2 inhibitors AAZ and benzolamide on the growth of the C. albicans Nce103 deletion mutant expressing CAN2 (RH1). RH1 was spotted onto YNB agar at 1 × 10^5, 1 × 10^4, and 1 × 10^3 cells/ml in the presence of (i) 4% DMSO and 3 mM benzolamide for 96 h or (ii) 4% DMSO and 3 mM AAZ for 72 h.

Figure 4.
Fig. 4. Can2 complex with the product analog acetate. (a) Overall structure of the Can2/acetate complex (monomer C, gray) overlaid with the structure of uncomplexed Can2 (blue). The Zn^2+ ions are shown as orange spheres. (b) Overlay of the active sites of the Can2/acetate complex and the Can2 structure with a bound water molecule (blue: Can2/water structure; aquamarine: Can2/acetate monomer A; cyan: Can2/acetate monomer B; gray: Can2/acetate monomer C). (c) Modeling of a Can2/benzolamide complex. Residues coordinating the active-site ion, restricting the space available to the inhibitor, or reachable for the inhibitor moieties are labeled.

The above figures are reprinted by permission from Elsevier: J Mol Biol (2009, 385, 1207-1220) copyright 2009.

Figures were selected by an automated process.