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Targeting a surface cavity of 1-antitrypsin to prevent conformational disease

Targeting a surface cavity of 1-antitrypsin to prevent conformational disease
Targeting a surface cavity of 1-antitrypsin to prevent conformational disease
Conformational diseases are caused by a structural rearrangement within a protein that results in aberrant intermolecular linkage and tissue deposition. This is typified by the polymers that form with the Z deficiency variant of alpha(1)-antitrypsin (Glu-342-->Lys). These polymers are retained within hepatocytes to form inclusions that are associated with hepatitis, cirrhosis, and hepatocellular carcinoma. We have assessed a surface hydrophobic cavity in alpha(1)-antitrypsin as a potential target for rational drug design in order to prevent polymer formation and the associated liver disease. The introduction of either Thr-114-->Phe or Gly-117-->Phe on strand 2 of beta-sheet A within this cavity significantly raised the melting temperature and retarded polymer formation. Conversely, Leu-100-->Phe on helix D accelerated polymer formation, but this effect was abrogated by the addition of Thr-114-->Phe. None of these mutations affected the inhibitory activity of alpha(1)-antitrypsin. The importance of these observations was underscored by the finding that the Thr-114-->Phe mutation reduced polymer formation and increased the secretion of Z alpha(1)-antitrypsin from a Xenopus oocyte expression system. Moreover cysteine mutants within the hydrophobic pocket were able to bind a range of fluorophores illustrating the accessibility of the cavity to external agents. These results demonstrate the importance of this cavity as a site for drug design to ameliorate polymerization and prevent the associated conformational disease.
0021-9258
33060-33066
Parfrey, Helen
4218ef5d-db8d-4383-af21-036efc3f0887
Mahadeva, Ravi
40ef2d46-d603-428b-ba4b-783d425ec1ae
Ravenhill, Neil A.
f621445e-342a-4430-8c83-591314fd8bdb
Zhou, Aiwu
b340863e-caa1-4967-a157-96e4bc73e268
Dafforn, Timothy R.
b6f6e4b8-8bea-4a68-95b3-6a4ee89a4026
Foreman, Richard C.
5e7796fe-6411-4495-aa90-5f17d5d67ffa
Lomas, David A.
0641855a-442c-44b1-9684-f3e3cfc97808
Parfrey, Helen
4218ef5d-db8d-4383-af21-036efc3f0887
Mahadeva, Ravi
40ef2d46-d603-428b-ba4b-783d425ec1ae
Ravenhill, Neil A.
f621445e-342a-4430-8c83-591314fd8bdb
Zhou, Aiwu
b340863e-caa1-4967-a157-96e4bc73e268
Dafforn, Timothy R.
b6f6e4b8-8bea-4a68-95b3-6a4ee89a4026
Foreman, Richard C.
5e7796fe-6411-4495-aa90-5f17d5d67ffa
Lomas, David A.
0641855a-442c-44b1-9684-f3e3cfc97808

Parfrey, Helen, Mahadeva, Ravi, Ravenhill, Neil A., Zhou, Aiwu, Dafforn, Timothy R., Foreman, Richard C. and Lomas, David A. (2003) Targeting a surface cavity of 1-antitrypsin to prevent conformational disease. The Journal of Biological Chemistry, 278 (35), 33060-33066. (doi:10.1074/jbc.M302646200).

Record type: Article

Abstract

Conformational diseases are caused by a structural rearrangement within a protein that results in aberrant intermolecular linkage and tissue deposition. This is typified by the polymers that form with the Z deficiency variant of alpha(1)-antitrypsin (Glu-342-->Lys). These polymers are retained within hepatocytes to form inclusions that are associated with hepatitis, cirrhosis, and hepatocellular carcinoma. We have assessed a surface hydrophobic cavity in alpha(1)-antitrypsin as a potential target for rational drug design in order to prevent polymer formation and the associated liver disease. The introduction of either Thr-114-->Phe or Gly-117-->Phe on strand 2 of beta-sheet A within this cavity significantly raised the melting temperature and retarded polymer formation. Conversely, Leu-100-->Phe on helix D accelerated polymer formation, but this effect was abrogated by the addition of Thr-114-->Phe. None of these mutations affected the inhibitory activity of alpha(1)-antitrypsin. The importance of these observations was underscored by the finding that the Thr-114-->Phe mutation reduced polymer formation and increased the secretion of Z alpha(1)-antitrypsin from a Xenopus oocyte expression system. Moreover cysteine mutants within the hydrophobic pocket were able to bind a range of fluorophores illustrating the accessibility of the cavity to external agents. These results demonstrate the importance of this cavity as a site for drug design to ameliorate polymerization and prevent the associated conformational disease.

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More information

Submitted date: 14 March 2003
Published date: 29 August 2003

Identifiers

Local EPrints ID: 55721
URI: http://eprints.soton.ac.uk/id/eprint/55721
ISSN: 0021-9258
PURE UUID: f50d74f3-70b7-4a09-8f6e-1e38f2bb3ab6

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Date deposited: 05 Aug 2008
Last modified: 15 Mar 2024 10:57

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Contributors

Author: Helen Parfrey
Author: Ravi Mahadeva
Author: Neil A. Ravenhill
Author: Aiwu Zhou
Author: Timothy R. Dafforn
Author: Richard C. Foreman
Author: David A. Lomas

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