Structural comparison of protiated, H/D-exchanged and deuterated human carbonic anhydrase IX
(2019) In Acta Crystallographica Section D: Structural Biology 75(Pt 10). p.895-903- Abstract
Human carbonic anhydrase IX (CA IX) expression is upregulated in hypoxic solid tumours, promoting cell survival and metastasis. This observation has made CA IX a target for the development of CA isoform-selective inhibitors. To enable structural studies of CA IX-inhibitor complexes using X-ray and neutron crystallography, a CA IX surface variant (CA IXSV; the catalytic domain with six surface amino-acid substitutions) has been developed that can be routinely crystallized. Here, the preparation of protiated (H/H), H/D-exchanged (H/D) and deuterated (D/D) CA IXSV for crystallographic studies and their structural comparison are described. Four CA IXSV X-ray crystal structures are compared: two H/H crystal forms, an H/D crystal form and a... (More)
Human carbonic anhydrase IX (CA IX) expression is upregulated in hypoxic solid tumours, promoting cell survival and metastasis. This observation has made CA IX a target for the development of CA isoform-selective inhibitors. To enable structural studies of CA IX-inhibitor complexes using X-ray and neutron crystallography, a CA IX surface variant (CA IXSV; the catalytic domain with six surface amino-acid substitutions) has been developed that can be routinely crystallized. Here, the preparation of protiated (H/H), H/D-exchanged (H/D) and deuterated (D/D) CA IXSV for crystallographic studies and their structural comparison are described. Four CA IXSV X-ray crystal structures are compared: two H/H crystal forms, an H/D crystal form and a D/D crystal form. The overall active-site organization in each version is essentially the same, with only minor positional changes in active-site solvent, which may be owing to deuteration and/or resolution differences. Analysis of the crystal contacts and packing reveals different arrangements of CA IXSV compared with previous reports. To our knowledge, this is the first report comparing three different deuterium-labelled crystal structures of the same protein, marking an important step in validating the active-site structure of CA IXSV for neutron protein crystallography.
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- author
- Koruza, K LU ; Lafumat, B LU ; Nyblom, M LU ; Mahon, B P ; Knecht, W LU ; McKenna, R and Fisher, S Z LU
- organization
- publishing date
- 2019-10-01
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Antigens, Neoplasm/chemistry, Carbonic Anhydrase IX/chemistry, Catalytic Domain, Cloning, Molecular, Crystallography, X-Ray/methods, Deuterium, Escherichia coli/genetics, Humans, Models, Molecular
- in
- Acta Crystallographica Section D: Structural Biology
- volume
- 75
- issue
- Pt 10
- pages
- 9 pages
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- pmid:31588921
- scopus:85072919114
- ISSN
- 2059-7983
- DOI
- 10.1107/S2059798319010027
- language
- English
- LU publication?
- yes
- additional info
- open access.
- id
- 3b68a104-156d-4e1b-9641-8f5c2263760f
- date added to LUP
- 2020-07-09 16:33:31
- date last changed
- 2024-09-19 02:32:29
@article{3b68a104-156d-4e1b-9641-8f5c2263760f, abstract = {{<p>Human carbonic anhydrase IX (CA IX) expression is upregulated in hypoxic solid tumours, promoting cell survival and metastasis. This observation has made CA IX a target for the development of CA isoform-selective inhibitors. To enable structural studies of CA IX-inhibitor complexes using X-ray and neutron crystallography, a CA IX surface variant (CA IXSV; the catalytic domain with six surface amino-acid substitutions) has been developed that can be routinely crystallized. Here, the preparation of protiated (H/H), H/D-exchanged (H/D) and deuterated (D/D) CA IXSV for crystallographic studies and their structural comparison are described. Four CA IXSV X-ray crystal structures are compared: two H/H crystal forms, an H/D crystal form and a D/D crystal form. The overall active-site organization in each version is essentially the same, with only minor positional changes in active-site solvent, which may be owing to deuteration and/or resolution differences. Analysis of the crystal contacts and packing reveals different arrangements of CA IXSV compared with previous reports. To our knowledge, this is the first report comparing three different deuterium-labelled crystal structures of the same protein, marking an important step in validating the active-site structure of CA IXSV for neutron protein crystallography.</p>}}, author = {{Koruza, K and Lafumat, B and Nyblom, M and Mahon, B P and Knecht, W and McKenna, R and Fisher, S Z}}, issn = {{2059-7983}}, keywords = {{Antigens, Neoplasm/chemistry; Carbonic Anhydrase IX/chemistry; Catalytic Domain; Cloning, Molecular; Crystallography, X-Ray/methods; Deuterium; Escherichia coli/genetics; Humans; Models, Molecular}}, language = {{eng}}, month = {{10}}, number = {{Pt 10}}, pages = {{895--903}}, publisher = {{John Wiley & Sons Inc.}}, series = {{Acta Crystallographica Section D: Structural Biology}}, title = {{Structural comparison of protiated, H/D-exchanged and deuterated human carbonic anhydrase IX}}, url = {{http://dx.doi.org/10.1107/S2059798319010027}}, doi = {{10.1107/S2059798319010027}}, volume = {{75}}, year = {{2019}}, }