skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Carbon dioxide "trapped" in a β-carbonic anhydrase

Abstract

Carbonic anhydrases (CAs) are enzymes that catalyze the hydration/ dehydration of CO2/HCO3- with rates approaching diffusion-controlled limits (kcat/KM ~ 108 M–1s–1). Here, this family of enzymes has evolved disparate protein folds that all perform the same reaction at near catalytic perfection. Presented here is a structural study of a beta-CA (psCA3) expressed in Pseudomonas aeruginosa, in complex with CO2, using pressurized cryocooled crystallography. The structure has been refined to 1.6 angstrom resolution with Rcryst and Rfree values of 17.3 and 19.9%, respectively, and is compared with the α-CA, human CA isoform II (hCA II), the only other CA to have CO2, captured in its active site. Despite the lack of structural similarity between psCA3 and hCA II, the CO2, binding orientation relative to the zinc-bound solvent is identical. In addition, a second CO2, binding site was located at the dimer interface of psCA3. Interestingly, all β-CAs function as dirners or higher-order oligomeric states, and the CO2, bound at the interface may contribute to the allosteric nature of this family of enzymes or may be a convenient alternative binding site as this pocket has been previously shown to be a promiscuous site for a variety of ligands, including bicarbonate, sulfate, andmore » phosphate ions.« less

Authors:
 [1];  [2];  [3];  [2];  [3]
  1. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  2. Cornell Univ., Ithaca, NY (United States)
  3. Univ. of Florida, Gainesville, FL (United States)
Publication Date:
Research Org.:
Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1287017
Grant/Contract Number:  
AC05-00OR22725
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Biochemistry
Additional Journal Information:
Journal Volume: 54; Journal Issue: 43; Journal ID: ISSN 0006-2960
Publisher:
American Chemical Society (ACS)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; β-carbonic anhydrase; Pseudomonas aeruginosa; CO2 binding

Citation Formats

Aggarwal, Mayank, Chua, Teck Khiang, Pinard, Melissa A., Szebenyi, Doletha M., and McKenna, Robert. Carbon dioxide "trapped" in a β-carbonic anhydrase. United States: N. p., 2015. Web. doi:10.1021/acs.biochem.5b00987.
Aggarwal, Mayank, Chua, Teck Khiang, Pinard, Melissa A., Szebenyi, Doletha M., & McKenna, Robert. Carbon dioxide "trapped" in a β-carbonic anhydrase. United States. https://doi.org/10.1021/acs.biochem.5b00987
Aggarwal, Mayank, Chua, Teck Khiang, Pinard, Melissa A., Szebenyi, Doletha M., and McKenna, Robert. 2015. "Carbon dioxide "trapped" in a β-carbonic anhydrase". United States. https://doi.org/10.1021/acs.biochem.5b00987. https://www.osti.gov/servlets/purl/1287017.
@article{osti_1287017,
title = {Carbon dioxide "trapped" in a β-carbonic anhydrase},
author = {Aggarwal, Mayank and Chua, Teck Khiang and Pinard, Melissa A. and Szebenyi, Doletha M. and McKenna, Robert},
abstractNote = {Carbonic anhydrases (CAs) are enzymes that catalyze the hydration/ dehydration of CO2/HCO3- with rates approaching diffusion-controlled limits (kcat/KM ~ 108 M–1s–1). Here, this family of enzymes has evolved disparate protein folds that all perform the same reaction at near catalytic perfection. Presented here is a structural study of a beta-CA (psCA3) expressed in Pseudomonas aeruginosa, in complex with CO2, using pressurized cryocooled crystallography. The structure has been refined to 1.6 angstrom resolution with Rcryst and Rfree values of 17.3 and 19.9%, respectively, and is compared with the α-CA, human CA isoform II (hCA II), the only other CA to have CO2, captured in its active site. Despite the lack of structural similarity between psCA3 and hCA II, the CO2, binding orientation relative to the zinc-bound solvent is identical. In addition, a second CO2, binding site was located at the dimer interface of psCA3. Interestingly, all β-CAs function as dirners or higher-order oligomeric states, and the CO2, bound at the interface may contribute to the allosteric nature of this family of enzymes or may be a convenient alternative binding site as this pocket has been previously shown to be a promiscuous site for a variety of ligands, including bicarbonate, sulfate, and phosphate ions.},
doi = {10.1021/acs.biochem.5b00987},
url = {https://www.osti.gov/biblio/1287017}, journal = {Biochemistry},
issn = {0006-2960},
number = 43,
volume = 54,
place = {United States},
year = {Mon Oct 12 00:00:00 EDT 2015},
month = {Mon Oct 12 00:00:00 EDT 2015}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Citation Metrics:
Cited by: 17 works
Citation information provided by
Web of Science

Save / Share:

Works referenced in this record:

Pseudomonas aeruginosa isoles de patients immunodéprimés : résistance aux antibiotiques, sérotypage et typage moléculaire
journal, November 2005


Why Is Carbonic Anhydrase Essential to Escherichia coli?
journal, November 2003


Possible Roles for His 208 in the Active-Site Region of Chloroplast Carbonic Anhydrase fromPisum sativum
journal, January 1999


Three functional β-carbonic anhydrases in Pseudomonas aeruginosa PAO1: role in survival in ambient air
journal, August 2013


Crystal structure of E. coli β-carbonic anhydrase, an enzyme with an unusual pH-dependent activity
journal, May 2001


Carbonic anhydrase is an ancient enzyme widespread in prokaryotes
journal, December 1999


Structural study of X-ray induced activation of carbonic anhydrase
journal, June 2009


Pseudomonas aeruginosa Outbreak in a Burn Unit: Role of Antimicrobials in the Emergence of Multiply Resistant Strains
journal, August 1994


High-pressure cooling of protein crystals without cryoprotectants
journal, June 2005


A gene homologous to chloroplast carbonic anhydrase (icfA) is essential to photosynthetic carbon dioxide fixation by Synechococcus PCC7942.
journal, May 1992


Coot model-building tools for molecular graphics
journal, November 2004


Carbonic Anhydrase Is Essential for Streptococcus pneumoniae Growth in Environmental Ambient Air
journal, August 2010


Pseudomonas aeruginosa: resistance and therapeutic options at the turn of the new millennium
journal, June 2007


Kinetic characterization of wild-type and proton transfer-impaired variants of β-carbonic anhydrase from Arabidopsis thaliana
journal, August 2002


Identification of a Novel Noncatalytic Bicarbonate Binding Site in Eubacterial β-Carbonic Anhydrase
journal, March 2006


Carbonic Anhydrase Is Essential for Growth of Ralstonia eutropha at Ambient CO2 Concentrations
journal, September 2002


Roles of α and β Carbonic Anhydrases of Helicobacter pylori in the Urease-Dependent Response to Acidity and in Colonization of the Murine Gastric Mucosa
journal, February 2008


Structure and inhibition studies of a type II beta-carbonic anhydrase psCA3 from Pseudomonas aeruginosa
journal, August 2015


PHENIX: a comprehensive Python-based system for macromolecular structure solution
journal, January 2010


Prokaryotic carbonic anhydrases
journal, October 2000


Allosteric Site Variants of Haemophilus influenzae β-Carbonic Anhydrase
journal, June 2009


Structural annotation of human carbonic anhydrases
journal, November 2012


Structure and catalytic mechanism of the β-carbonic anhydrases
journal, February 2010


A high-pressure cryocooling method for protein crystals and biological samples with reduced background X-ray scatter
journal, December 2012


Roles of the Conserved Aspartate and Arginine in the Catalytic Mechanism of an Archaeal β-Class Carbonic Anhydrase
journal, August 2002


A physiological role for cyanate-induced carbonic anhydrase in Escherichia coli
journal, March 1993


A gene homologous to ?-type carbonic anhydrase is essential for the growth of Corynebacterium glutamicum under atmospheric conditions
journal, May 2004


Inference of Macromolecular Assemblies from Crystalline State
journal, September 2007


Entrapment of Carbon Dioxide in the Active Site of Carbonic Anhydrase II
journal, September 2008


[20] Processing of X-ray diffraction data collected in oscillation mode
book, January 1997


Structural insight into activity enhancement and inhibition of H64A carbonic anhydrase II by imidazoles
journal, February 2014


Fluorescence-Based Isolation of Bacterial Genes Expressed Within Host Cells
journal, September 1997


Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX
journal, September 2009


Carbonic anhydrase in Escherichia coli. A product of the cyn operon.
journal, February 1992


Works referencing / citing this record:

Kinetic study of catalytic CO 2 hydration by metal-substituted biomimetic carbonic anhydrase model complexes
journal, August 2019


Structure and function of carbonic anhydrases
journal, January 1977


Structure and Function of Carbonic Anhydrases
book, January 1981