The Reactivity and Oxidation Pathway of Cysteine 232 in Recombinant Human α1-Antitrypsin
Oxidative damage to the sulfur-containing amino acids, methionine and cysteine, is a major concern in biotechnology and medicine. α1-Antitrypsin, which is a metastable and conformationally flexible protein that belongs to the serpin family of protease inhibitors, contains nine methionines and a sing...
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| مؤلفون آخرون: | , |
| التنسيق: | article |
| منشور في: |
2002
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| الوصول للمادة أونلاين: | https://hdl.handle.net/11073/25757 |
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| _version_ | 1864513432844238848 |
|---|---|
| author | Griffiths, Steven |
| author2 | King, Jonathan Cooney, Charles L. |
| author2_role | author author |
| author_facet | Griffiths, Steven King, Jonathan Cooney, Charles L. |
| author_role | author |
| dc.creator.none.fl_str_mv | Griffiths, Steven King, Jonathan Cooney, Charles L. |
| dc.date.none.fl_str_mv | 2002-07-12 2024-12-31T08:50:42Z 2024-12-31T08:50:42Z |
| dc.format.none.fl_str_mv | application/pdf |
| dc.identifier.none.fl_str_mv | Griffiths, S. W., King, J., & Cooney, C. L. (2002). The Reactivity and Oxidation Pathway of Cysteine 232 in Recombinant Human α1-Antitrypsin. In Journal of Biological Chemistry (Vol. 277, Issue 28, pp. 25486–25492). Elsevier BV. https://doi.org/10.1074/jbc.m203089200 0021-9258 https://hdl.handle.net/11073/25757 10.1074/jbc.m203089200 |
| dc.language.none.fl_str_mv | en |
| dc.publisher.none.fl_str_mv | The American Society for Biochemistry and Molecular Biology |
| dc.relation.none.fl_str_mv | https://doi.org/10.1074/jbc.m203089200 |
| dc.rights.none.fl_str_mv | Attribution 4.0 International http://creativecommons.org/licenses/by/4.0/ |
| dc.title.none.fl_str_mv | The Reactivity and Oxidation Pathway of Cysteine 232 in Recombinant Human α1-Antitrypsin |
| dc.type.none.fl_str_mv | Peer-Reviewed Published version info:eu-repo/semantics/publishedVersion info:eu-repo/semantics/article |
| description | Oxidative damage to the sulfur-containing amino acids, methionine and cysteine, is a major concern in biotechnology and medicine. α1-Antitrypsin, which is a metastable and conformationally flexible protein that belongs to the serpin family of protease inhibitors, contains nine methionines and a single cysteine in its primary sequence. Although it is known that methionine oxidation in the protein active site results in a loss of biological activity, there is little specific knowledge regarding the reactivity of its unpaired thiol, Cys-232. In this study, the thiol-modifying reagent NBD-Cl (7-chloro-4-nitrobenz-2-oxa-1,3-diazole) was used to label peroxide-modified α1-antitrypsin and demonstrate that the Cys-232in vitro oxidation pathway begins with a stable sulfenic acid intermediate and is followed by the formation of sulfinic and cysteic acid in successive steps. pH-dependent reactivity with hydrogen peroxide showed that Cys-232 has a pK a of 6.86 ± 0.05, a value that is more than 1.5 pH units lower than that of a typical protein thiol. pH-induced conformational changes in the region surrounding Cys-232 were also examined and indicate that mildly acidic conditions induce a conformation that enhances Cys-232 reactivity. In summary, this work provides new insights into α1-antitrypsin reactivity in oxidizing environments and shows that a unique structural environment renders its unpaired thiol, Cys-232, its most reactive amino acid. |
| format | article |
| id | aus_7155bbd297f29f11200702ee83a04700 |
| identifier_str_mv | Griffiths, S. W., King, J., & Cooney, C. L. (2002). The Reactivity and Oxidation Pathway of Cysteine 232 in Recombinant Human α1-Antitrypsin. In Journal of Biological Chemistry (Vol. 277, Issue 28, pp. 25486–25492). Elsevier BV. https://doi.org/10.1074/jbc.m203089200 0021-9258 10.1074/jbc.m203089200 |
| language_invalid_str_mv | en |
| network_acronym_str | aus |
| network_name_str | aus |
| oai_identifier_str | oai:repository.aus.edu:11073/25757 |
| publishDate | 2002 |
| publisher.none.fl_str_mv | The American Society for Biochemistry and Molecular Biology |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | Attribution 4.0 International http://creativecommons.org/licenses/by/4.0/ |
| spelling | The Reactivity and Oxidation Pathway of Cysteine 232 in Recombinant Human α1-AntitrypsinGriffiths, StevenKing, JonathanCooney, Charles L.Oxidative damage to the sulfur-containing amino acids, methionine and cysteine, is a major concern in biotechnology and medicine. α1-Antitrypsin, which is a metastable and conformationally flexible protein that belongs to the serpin family of protease inhibitors, contains nine methionines and a single cysteine in its primary sequence. Although it is known that methionine oxidation in the protein active site results in a loss of biological activity, there is little specific knowledge regarding the reactivity of its unpaired thiol, Cys-232. In this study, the thiol-modifying reagent NBD-Cl (7-chloro-4-nitrobenz-2-oxa-1,3-diazole) was used to label peroxide-modified α1-antitrypsin and demonstrate that the Cys-232in vitro oxidation pathway begins with a stable sulfenic acid intermediate and is followed by the formation of sulfinic and cysteic acid in successive steps. pH-dependent reactivity with hydrogen peroxide showed that Cys-232 has a pK a of 6.86 ± 0.05, a value that is more than 1.5 pH units lower than that of a typical protein thiol. pH-induced conformational changes in the region surrounding Cys-232 were also examined and indicate that mildly acidic conditions induce a conformation that enhances Cys-232 reactivity. In summary, this work provides new insights into α1-antitrypsin reactivity in oxidizing environments and shows that a unique structural environment renders its unpaired thiol, Cys-232, its most reactive amino acid.The American Society for Biochemistry and Molecular Biology2024-12-31T08:50:42Z2024-12-31T08:50:42Z2002-07-12Peer-ReviewedPublished versioninfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfGriffiths, S. W., King, J., & Cooney, C. L. (2002). The Reactivity and Oxidation Pathway of Cysteine 232 in Recombinant Human α1-Antitrypsin. In Journal of Biological Chemistry (Vol. 277, Issue 28, pp. 25486–25492). Elsevier BV. https://doi.org/10.1074/jbc.m2030892000021-9258https://hdl.handle.net/11073/2575710.1074/jbc.m203089200enhttps://doi.org/10.1074/jbc.m203089200Attribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/oai:repository.aus.edu:11073/257572024-12-31T15:01:30Z |
| spellingShingle | The Reactivity and Oxidation Pathway of Cysteine 232 in Recombinant Human α1-Antitrypsin Griffiths, Steven |
| status_str | publishedVersion |
| title | The Reactivity and Oxidation Pathway of Cysteine 232 in Recombinant Human α1-Antitrypsin |
| title_full | The Reactivity and Oxidation Pathway of Cysteine 232 in Recombinant Human α1-Antitrypsin |
| title_fullStr | The Reactivity and Oxidation Pathway of Cysteine 232 in Recombinant Human α1-Antitrypsin |
| title_full_unstemmed | The Reactivity and Oxidation Pathway of Cysteine 232 in Recombinant Human α1-Antitrypsin |
| title_short | The Reactivity and Oxidation Pathway of Cysteine 232 in Recombinant Human α1-Antitrypsin |
| title_sort | The Reactivity and Oxidation Pathway of Cysteine 232 in Recombinant Human α1-Antitrypsin |
| url | https://hdl.handle.net/11073/25757 |