Structure and Inhibition of the CO2-Sensing Carbonic Anhydrase Can2 from the Pathogenic Fungus Cryptococcus neoformans

被引:164
作者
Schlicker, Christine [1 ]
Hall, Rebecca A. [2 ]
Vullo, Daniela [3 ]
Middelhaufe, Sabine [1 ]
Gertz, Melanie [1 ]
Supuran, Claudiu T. [3 ]
Muehlschlegel, Fritz A. [2 ]
Steegborn, Clemens [1 ]
机构
[1] Ruhr Univ Bochum, Dept Physiol Chem, D-44801 Bochum, Germany
[2] Univ Kent, Dept Biosci, Canterbury CT2 7NJ, Kent, England
[3] Univ Florence, Lab Chim Bioinorgan, I-50019 Florence, Italy
基金
英国医学研究理事会;
关键词
beta-class carbonic anhydrase; Cryptococcus neoformans; crystal structure; inhibition; sulfonamide; PH-DEPENDENT ACTIVITY; BETA-CLASS; GAMMA-CLASS; MYCOBACTERIUM-TUBERCULOSIS; MACROMOLECULAR STRUCTURES; THERAPEUTIC APPLICATIONS; MALARIA PARASITES; CRYSTAL-STRUCTURE; ANCIENT ENZYME; ACTIVE-SITE;
D O I
10.1016/j.jmb.2008.11.037
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In the pathogenic fungus Cryptococcus neoformans, a CO2-sensing system is essential for survival in the natural environment (similar to 0.03% CO2) and mediates the switch to virulent growth in the human host (similar to 5% CO2). 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. (C) 2008 Elsevier Ltd. All rights resented.
引用
收藏
页码:1207 / 1220
页数:14
相关论文
共 46 条
[31]   PRODRG:: a tool for high-throughput crystallography of protein-ligand complexes [J].
Schüttelkopf, AW ;
van Aalten, DMF .
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2004, 60 :1355-1363
[32]   SHELXL: High-resolution refinement [J].
Sheldrick, GM ;
Schneider, TR .
MACROMOLECULAR CRYSTALLOGRAPHY, PT B, 1997, 277 :319-343
[33]   Carbonic anhydrase is an ancient enzyme widespread in prokaryotes [J].
Smith, KS ;
Jakubzick, C ;
Whittam, TS ;
Ferry, JG .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (26) :15184-15189
[34]   Structural and kinetic characterization of an archaeal β-class carbonic anhydrase [J].
Smith, KS ;
Cosper, NJ ;
Stalhandske, C ;
Scott, RA ;
Ferry, JG .
JOURNAL OF BACTERIOLOGY, 2000, 182 (23) :6605-6613
[35]   A novel evolutionary lineage of carbonic anhydrase (ε class) is a component of the carboxysome shell [J].
So, AKC ;
Espie, GS ;
Williams, EB ;
Shively, JM ;
Heinhorst, S ;
Cannon, GC .
JOURNAL OF BACTERIOLOGY, 2004, 186 (03) :623-630
[36]   Crystal structure of the "cab"-type β class carbonic anhydrase from the archaeon Methanobacterium thermoautotrophicum [J].
Strop, P ;
Smith, KS ;
Iverson, TM ;
Ferry, JG ;
Rees, DC .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (13) :10299-10305
[37]   Carbonic anhydrases - An overview [J].
Supuran, Claudiu T. .
CURRENT PHARMACEUTICAL DESIGN, 2008, 14 (07) :603-614
[38]   Carbonic anhydrases: novel therapeutic applications for inhibitors and activators [J].
Supuran, Claudiu T. .
NATURE REVIEWS DRUG DISCOVERY, 2008, 7 (02) :168-181
[39]   Carbonic anhydrase inhibitors [J].
Supuran, CT ;
Scozzafava, A ;
Casini, A .
MEDICINAL RESEARCH REVIEWS, 2003, 23 (02) :146-189
[40]   Carbonic anhydrase activators: The first X-ray crystallographic study of an adduct of isoform I [J].
Temperini, Claudia ;
Scozzafava, Andrea ;
Supuran, Claudiu T. .
BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, 2006, 16 (19) :5152-5156