Structural and kinetic characterization of active-site histidine as a proton shuttle in catalysis by human carbonic anhydrase II

Zoë Fisher, Jose A. Hernandez Prada, Chingkuang Tu, David Duda, Craig Yoshioka, Haiqian An, Lakshmanan Govindasamy, David N. Silverman, Robert McKenna

Research output: Contribution to journalArticle

108 Citations (Scopus)

Abstract

In the catalysis of the hydration of carbon dioxide and dehydration of bicarbonate by human carbonic anhydrase II (HCA II), a histidine residue (His64) shuttles protons between the zinc-bound solvent molecule and the bulk solution. To evaluate the effect of the position of the shuttle histidine and pH on proton shuttling, we have examined the catalysis and crystal structures of wild-type HCA II and two double mutants: H64A/N62H and H64A/N67H HCA II. His62 and His67 both have their side chains extending into the active-site cavity with distances from the zinc approximately equivalent to that of His64. Crystal structures were determined at pH 5.1-10.0, and the catalysis of the exchange of 18O between CO2 and water was assessed by mass spectrometry. Efficient proton shuttle exceeding a rate of 105 s -1 was observed for histidine at positions 64 and 67; in contrast, relatively inefficient proton transfer at a rate near 103 s -1 was observed for His62. The observation, in the crystal structures, of a completed hydrogen-bonded water chain between the histidine shuttle residue and the zinc-bound solvent does not appear to be required for efficient proton transfer. The data suggest that the number of intervening water molecules between the donor and acceptor supporting efficient proton transfer in HCA II is important, and furthermore suggest that a water bridge consisting of two intervening water molecules is consistent with efficient proton transfer.

Original languageEnglish (US)
Pages (from-to)1097-1105
Number of pages9
JournalBiochemistry
Volume44
Issue number4
DOIs
StatePublished - Feb 1 2005

Fingerprint

Carbonic Anhydrase II
Catalysis
Histidine
Proton transfer
Protons
Catalytic Domain
Kinetics
Water
Zinc
Crystal structure
Molecules
Bicarbonates
Dehydration
Carbon Dioxide
Hydration
Mass spectrometry
Hydrogen
Ion exchange
Mass Spectrometry
Observation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Structural and kinetic characterization of active-site histidine as a proton shuttle in catalysis by human carbonic anhydrase II. / Fisher, Zoë; Hernandez Prada, Jose A.; Tu, Chingkuang; Duda, David; Yoshioka, Craig; An, Haiqian; Govindasamy, Lakshmanan; Silverman, David N.; McKenna, Robert.

In: Biochemistry, Vol. 44, No. 4, 01.02.2005, p. 1097-1105.

Research output: Contribution to journalArticle

Fisher, Z, Hernandez Prada, JA, Tu, C, Duda, D, Yoshioka, C, An, H, Govindasamy, L, Silverman, DN & McKenna, R 2005, 'Structural and kinetic characterization of active-site histidine as a proton shuttle in catalysis by human carbonic anhydrase II', Biochemistry, vol. 44, no. 4, pp. 1097-1105. https://doi.org/10.1021/bi0480279
Fisher, Zoë ; Hernandez Prada, Jose A. ; Tu, Chingkuang ; Duda, David ; Yoshioka, Craig ; An, Haiqian ; Govindasamy, Lakshmanan ; Silverman, David N. ; McKenna, Robert. / Structural and kinetic characterization of active-site histidine as a proton shuttle in catalysis by human carbonic anhydrase II. In: Biochemistry. 2005 ; Vol. 44, No. 4. pp. 1097-1105.
@article{a9a37b05aab44f2bb314a82b4780634a,
title = "Structural and kinetic characterization of active-site histidine as a proton shuttle in catalysis by human carbonic anhydrase II",
abstract = "In the catalysis of the hydration of carbon dioxide and dehydration of bicarbonate by human carbonic anhydrase II (HCA II), a histidine residue (His64) shuttles protons between the zinc-bound solvent molecule and the bulk solution. To evaluate the effect of the position of the shuttle histidine and pH on proton shuttling, we have examined the catalysis and crystal structures of wild-type HCA II and two double mutants: H64A/N62H and H64A/N67H HCA II. His62 and His67 both have their side chains extending into the active-site cavity with distances from the zinc approximately equivalent to that of His64. Crystal structures were determined at pH 5.1-10.0, and the catalysis of the exchange of 18O between CO2 and water was assessed by mass spectrometry. Efficient proton shuttle exceeding a rate of 105 s -1 was observed for histidine at positions 64 and 67; in contrast, relatively inefficient proton transfer at a rate near 103 s -1 was observed for His62. The observation, in the crystal structures, of a completed hydrogen-bonded water chain between the histidine shuttle residue and the zinc-bound solvent does not appear to be required for efficient proton transfer. The data suggest that the number of intervening water molecules between the donor and acceptor supporting efficient proton transfer in HCA II is important, and furthermore suggest that a water bridge consisting of two intervening water molecules is consistent with efficient proton transfer.",
author = "Zo{\"e} Fisher and {Hernandez Prada}, {Jose A.} and Chingkuang Tu and David Duda and Craig Yoshioka and Haiqian An and Lakshmanan Govindasamy and Silverman, {David N.} and Robert McKenna",
year = "2005",
month = "2",
day = "1",
doi = "10.1021/bi0480279",
language = "English (US)",
volume = "44",
pages = "1097--1105",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "4",

}

TY - JOUR

T1 - Structural and kinetic characterization of active-site histidine as a proton shuttle in catalysis by human carbonic anhydrase II

AU - Fisher, Zoë

AU - Hernandez Prada, Jose A.

AU - Tu, Chingkuang

AU - Duda, David

AU - Yoshioka, Craig

AU - An, Haiqian

AU - Govindasamy, Lakshmanan

AU - Silverman, David N.

AU - McKenna, Robert

PY - 2005/2/1

Y1 - 2005/2/1

N2 - In the catalysis of the hydration of carbon dioxide and dehydration of bicarbonate by human carbonic anhydrase II (HCA II), a histidine residue (His64) shuttles protons between the zinc-bound solvent molecule and the bulk solution. To evaluate the effect of the position of the shuttle histidine and pH on proton shuttling, we have examined the catalysis and crystal structures of wild-type HCA II and two double mutants: H64A/N62H and H64A/N67H HCA II. His62 and His67 both have their side chains extending into the active-site cavity with distances from the zinc approximately equivalent to that of His64. Crystal structures were determined at pH 5.1-10.0, and the catalysis of the exchange of 18O between CO2 and water was assessed by mass spectrometry. Efficient proton shuttle exceeding a rate of 105 s -1 was observed for histidine at positions 64 and 67; in contrast, relatively inefficient proton transfer at a rate near 103 s -1 was observed for His62. The observation, in the crystal structures, of a completed hydrogen-bonded water chain between the histidine shuttle residue and the zinc-bound solvent does not appear to be required for efficient proton transfer. The data suggest that the number of intervening water molecules between the donor and acceptor supporting efficient proton transfer in HCA II is important, and furthermore suggest that a water bridge consisting of two intervening water molecules is consistent with efficient proton transfer.

AB - In the catalysis of the hydration of carbon dioxide and dehydration of bicarbonate by human carbonic anhydrase II (HCA II), a histidine residue (His64) shuttles protons between the zinc-bound solvent molecule and the bulk solution. To evaluate the effect of the position of the shuttle histidine and pH on proton shuttling, we have examined the catalysis and crystal structures of wild-type HCA II and two double mutants: H64A/N62H and H64A/N67H HCA II. His62 and His67 both have their side chains extending into the active-site cavity with distances from the zinc approximately equivalent to that of His64. Crystal structures were determined at pH 5.1-10.0, and the catalysis of the exchange of 18O between CO2 and water was assessed by mass spectrometry. Efficient proton shuttle exceeding a rate of 105 s -1 was observed for histidine at positions 64 and 67; in contrast, relatively inefficient proton transfer at a rate near 103 s -1 was observed for His62. The observation, in the crystal structures, of a completed hydrogen-bonded water chain between the histidine shuttle residue and the zinc-bound solvent does not appear to be required for efficient proton transfer. The data suggest that the number of intervening water molecules between the donor and acceptor supporting efficient proton transfer in HCA II is important, and furthermore suggest that a water bridge consisting of two intervening water molecules is consistent with efficient proton transfer.

UR - http://www.scopus.com/inward/record.url?scp=13444269025&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=13444269025&partnerID=8YFLogxK

U2 - 10.1021/bi0480279

DO - 10.1021/bi0480279

M3 - Article

C2 - 15667203

AN - SCOPUS:13444269025

VL - 44

SP - 1097

EP - 1105

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 4

ER -