Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX

Vincenzo Alterio, Mika Hilvo, Anna Di Fiore, Claudiu T. Supuran, Peiwen Pan, Seppo Parkkila, Andrea Scaloni, Jaromir Pastorek, Silvia Pastorekova, Carlo Pedone, Andrea Scozzafava, Simona Maria Monti, Giuseppina De Simone

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Abstract

Carbonic anhydrase (CA) IX is a plasma membrane-associated member of the α-CA enzyme family, which is involved in solid tumor acidification. It is a marker of tumor hypoxia and a prognostic factor in several human cancers. An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. Here we report the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide. The structure reveals a typical α-CA fold, which significantly differs from the other CA isozymes when the protein quaternary structure is considered. Thus, two catalytic domains of CA IX associate to form a dimer, which is stabilized by the formation of an intermolecular disulfide bond. The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. A correlation between the three-dimensional structure and the physiological role of the enzyme is here suggested, based on the measurement of the pH profile of the catalytic activity for the physiological reaction, CO2 hydration to bicarbonate and protons. On the basis of the structural differences observed between CA IX and the other membrane-associated α-CAs, further prospects for the rational drug design of isozyme-specific CA inhibitors are proposed, given that inhibition of this enzyme shows antitumor activity both in vitro and in vivo.
Original languageEnglish
Pages (from-to)16233-16238
JournalProceedings of the National Academy of Sciences of the United States of America
Volume106
Issue number38
DOIs
Publication statusPublished - 2009
MoE publication typeA1 Journal article-refereed

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Carbonic Anhydrases
Tumors
Crystal structure
Cell membranes
Dimers
Isoenzymes
Enzymes
Carbonic Anhydrase Inhibitors
Acetazolamide
human CA9 protein
Acidification
Cell adhesion
Sulfonamides
Tumor Biomarkers
Bicarbonates
Disulfides
Hydration
Protons
Catalyst activity
Proteins

Cite this

Alterio, Vincenzo ; Hilvo, Mika ; Di Fiore, Anna ; Supuran, Claudiu T. ; Pan, Peiwen ; Parkkila, Seppo ; Scaloni, Andrea ; Pastorek, Jaromir ; Pastorekova, Silvia ; Pedone, Carlo ; Scozzafava, Andrea ; Monti, Simona Maria ; De Simone, Giuseppina. / Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX. In: Proceedings of the National Academy of Sciences of the United States of America. 2009 ; Vol. 106, No. 38. pp. 16233-16238.
@article{0dad461c1816487bbd35626b903bf8b7,
title = "Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX",
abstract = "Carbonic anhydrase (CA) IX is a plasma membrane-associated member of the α-CA enzyme family, which is involved in solid tumor acidification. It is a marker of tumor hypoxia and a prognostic factor in several human cancers. An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. Here we report the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide. The structure reveals a typical α-CA fold, which significantly differs from the other CA isozymes when the protein quaternary structure is considered. Thus, two catalytic domains of CA IX associate to form a dimer, which is stabilized by the formation of an intermolecular disulfide bond. The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. A correlation between the three-dimensional structure and the physiological role of the enzyme is here suggested, based on the measurement of the pH profile of the catalytic activity for the physiological reaction, CO2 hydration to bicarbonate and protons. On the basis of the structural differences observed between CA IX and the other membrane-associated α-CAs, further prospects for the rational drug design of isozyme-specific CA inhibitors are proposed, given that inhibition of this enzyme shows antitumor activity both in vitro and in vivo.",
author = "Vincenzo Alterio and Mika Hilvo and {Di Fiore}, Anna and Supuran, {Claudiu T.} and Peiwen Pan and Seppo Parkkila and Andrea Scaloni and Jaromir Pastorek and Silvia Pastorekova and Carlo Pedone and Andrea Scozzafava and Monti, {Simona Maria} and {De Simone}, Giuseppina",
year = "2009",
doi = "10.1073/pnas.0908301106",
language = "English",
volume = "106",
pages = "16233--16238",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
publisher = "National Academy of Sciences",
number = "38",

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Alterio, V, Hilvo, M, Di Fiore, A, Supuran, CT, Pan, P, Parkkila, S, Scaloni, A, Pastorek, J, Pastorekova, S, Pedone, C, Scozzafava, A, Monti, SM & De Simone, G 2009, 'Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX', Proceedings of the National Academy of Sciences of the United States of America, vol. 106, no. 38, pp. 16233-16238. https://doi.org/10.1073/pnas.0908301106

Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX. / Alterio, Vincenzo; Hilvo, Mika; Di Fiore, Anna; Supuran, Claudiu T.; Pan, Peiwen; Parkkila, Seppo; Scaloni, Andrea; Pastorek, Jaromir; Pastorekova, Silvia; Pedone, Carlo; Scozzafava, Andrea; Monti, Simona Maria; De Simone, Giuseppina.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 106, No. 38, 2009, p. 16233-16238.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX

AU - Alterio, Vincenzo

AU - Hilvo, Mika

AU - Di Fiore, Anna

AU - Supuran, Claudiu T.

AU - Pan, Peiwen

AU - Parkkila, Seppo

AU - Scaloni, Andrea

AU - Pastorek, Jaromir

AU - Pastorekova, Silvia

AU - Pedone, Carlo

AU - Scozzafava, Andrea

AU - Monti, Simona Maria

AU - De Simone, Giuseppina

PY - 2009

Y1 - 2009

N2 - Carbonic anhydrase (CA) IX is a plasma membrane-associated member of the α-CA enzyme family, which is involved in solid tumor acidification. It is a marker of tumor hypoxia and a prognostic factor in several human cancers. An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. Here we report the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide. The structure reveals a typical α-CA fold, which significantly differs from the other CA isozymes when the protein quaternary structure is considered. Thus, two catalytic domains of CA IX associate to form a dimer, which is stabilized by the formation of an intermolecular disulfide bond. The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. A correlation between the three-dimensional structure and the physiological role of the enzyme is here suggested, based on the measurement of the pH profile of the catalytic activity for the physiological reaction, CO2 hydration to bicarbonate and protons. On the basis of the structural differences observed between CA IX and the other membrane-associated α-CAs, further prospects for the rational drug design of isozyme-specific CA inhibitors are proposed, given that inhibition of this enzyme shows antitumor activity both in vitro and in vivo.

AB - Carbonic anhydrase (CA) IX is a plasma membrane-associated member of the α-CA enzyme family, which is involved in solid tumor acidification. It is a marker of tumor hypoxia and a prognostic factor in several human cancers. An aberrant increase in CA IX expression in chronic hypoxia and during development of various carcinomas contributes to tumorigenesis through at least two mechanisms: pH regulation and cell adhesion control. Here we report the X-ray structure of the catalytic domain of CA IX in complex with a classical, clinically used sulfonamide inhibitor, acetazolamide. The structure reveals a typical α-CA fold, which significantly differs from the other CA isozymes when the protein quaternary structure is considered. Thus, two catalytic domains of CA IX associate to form a dimer, which is stabilized by the formation of an intermolecular disulfide bond. The active site clefts and the PG domains are located on one face of the dimer, while the C-termini are located on the opposite face to facilitate protein anchoring to the cell membrane. A correlation between the three-dimensional structure and the physiological role of the enzyme is here suggested, based on the measurement of the pH profile of the catalytic activity for the physiological reaction, CO2 hydration to bicarbonate and protons. On the basis of the structural differences observed between CA IX and the other membrane-associated α-CAs, further prospects for the rational drug design of isozyme-specific CA inhibitors are proposed, given that inhibition of this enzyme shows antitumor activity both in vitro and in vivo.

U2 - 10.1073/pnas.0908301106

DO - 10.1073/pnas.0908301106

M3 - Article

VL - 106

SP - 16233

EP - 16238

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 38

ER -