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Title: A sucrose-binding site provides a lead towards an isoform-specific inhibitor of the cancer-associated enzyme carbonic anhydrase IX

Journal Article · · Acta Crystallographica. Section F, Structural Biology Communications
 [1];  [2];  [1];  [3];  [1]
  1. Univ. of Florida, Gainesville, FL (United States). College of Medicine, Dept. of Biochemistry and Molecular Biology
  2. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Division of Biology and Soft Matter
  3. Univ. of Florida, Gainesville, FL (United States). College of Medicine, Dept. of Pharmacology
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Human carbonic anhydrase (CA; EC 4.2.1.1) isoform IX (CA IX) is an extracellular zinc metalloenzyme that catalyzes the reversible hydration of CO2to HCO3$$-$$, thereby playing a role in pH regulation. The majority of normal functioning cells exhibit low-level expression of CA IX. However, in cancer cells CA IX is upregulated as a consequence of a metabolic transition known as the Warburg effect. The upregulation of CA IX for cancer progression has drawn interest in it being a potential therapeutic target. CA IX is a transmembrane protein, and its purification, yield and crystallization have proven challenging to structure-based drug design, whereas the closely related cytosolic soluble isoform CA II can be expressed and crystallized with ease. Therefore, we have utilized structural alignments and site-directed mutagenesis to engineer a CA II that mimics the active site of CA IX. In this paper, the X-ray crystal structure of this CA IX mimic in complex with sucrose is presented and has been refined to a resolution of 1.5 Å, anRcryst of 18.0% and anRfree of 21.2%. Finally, the binding of sucrose at the entrance to the active site of the CA IX mimic, and not CA II, in a non-inhibitory mechanism provides a novel carbohydrate moiety binding site that could be further exploited to design isoform-specific inhibitors of CA IX.

Research Organization:
Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
Sponsoring Organization:
USDOE
DOE Contract Number:
AC05-00OR22725; GM25154
OSTI ID:
1343505
Journal Information:
Acta Crystallographica. Section F, Structural Biology Communications, Vol. 71, Issue 10; ISSN 2053-230X
Publisher:
International Union of Crystallography
Country of Publication:
United States
Language:
English

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Figures / Tables (1)

Table 1(p. 11)table Table 1

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
60 APPLIED LIFE SCIENCES
alpha-carbonic anhydrase
CA IX mimic
isoform-specific drug design
sugar approach
carbonic anhydrase IX
sucrose