X-ray structure of Candida antarctica lipase A shows a novel lid structure and a likely mode of interfacial activation
In nature, lipases (EC 3.1.1.3) catalyze the hydrolysis of triglycerides to form glycerol and fatty acids. Under the appropriate conditions, the reaction is reversible, and so biotechnological applications commonly make use of their capacity for esterification as well as for hydrolysis of a wide var...
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Uppsala universitet, Strukturell molekylärbiologi
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ftuppsalauniv:oai:DiVA.org:uu-14221 2023-05-15T14:03:20+02:00 X-ray structure of Candida antarctica lipase A shows a novel lid structure and a likely mode of interfacial activation Ericsson, Daniel J. Kasrayan, Alex Johansson, Patrik Bergfors, Terese Sandström, Anders G. Bäckvall, Jan-Erling Mowbray, Sherry L. 2008 application/pdf http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-14221 https://doi.org/10.1016/j.jmb.2007.10.079 eng eng Uppsala universitet, Strukturell molekylärbiologi Department of Molecular Biology, Swedish University of Agricultural Sciences, Biomedical Center, Uppsala, Sweden Journal of Molecular Biology, 0022-2836, 2008, 376:1, s. 109-119 http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-14221 doi:10.1016/j.jmb.2007.10.079 PMID 18155238 ISI:000253181500011 info:eu-repo/semantics/openAccess lipase interfacial activation hydrolase X-ray structure substrate specificity Biochemistry and Molecular Biology Biokemi och molekylärbiologi Article in journal info:eu-repo/semantics/article text 2008 ftuppsalauniv https://doi.org/10.1016/j.jmb.2007.10.079 2023-02-23T21:45:48Z In nature, lipases (EC 3.1.1.3) catalyze the hydrolysis of triglycerides to form glycerol and fatty acids. Under the appropriate conditions, the reaction is reversible, and so biotechnological applications commonly make use of their capacity for esterification as well as for hydrolysis of a wide variety of compounds. In the present paper, we report the X-ray structure of lipase A from Candida antarctica, solved by single isomorphous replacement with anomalous scattering, and refined to 2.2-A resolution. The structure is the first from a novel family of lipases. Contrary to previous predictions, the fold includes a well-defined lid as well as a classic alpha/beta hydrolase domain. The catalytic triad is identified as Ser184, Asp334 and His366, which follow the sequential order considered to be characteristic of lipases; the serine lies within a typical nucleophilic elbow. Computer docking studies, as well as comparisons to related structures, place the carboxylate group of a fatty acid product near the serine nucleophile, with the long lipid tail closely following the path through the lid that is marked by a fortuitously bound molecule of polyethylene glycol. For an ester substrate to bind in an equivalent fashion, loop movements near Phe431 will be required, suggesting the primary focus of the conformational changes required for interfacial activation. Such movements will provide virtually unlimited access to solvent for the alcohol moiety of an ester substrate. The structure thus provides a basis for understanding the enzyme's preference for acyl moieties with long, straight tails, and for its highly promiscuous acceptance of widely different alcohol and amine moieties. An unconventional oxyanion hole is observed in the present structure, although the situation may change during interfacial activation. Article in Journal/Newspaper Antarc* Antarctica Uppsala University: Publications (DiVA) Journal of Molecular Biology 376 1 109 119 |
institution |
Open Polar |
collection |
Uppsala University: Publications (DiVA) |
op_collection_id |
ftuppsalauniv |
language |
English |
topic |
lipase interfacial activation hydrolase X-ray structure substrate specificity Biochemistry and Molecular Biology Biokemi och molekylärbiologi |
spellingShingle |
lipase interfacial activation hydrolase X-ray structure substrate specificity Biochemistry and Molecular Biology Biokemi och molekylärbiologi Ericsson, Daniel J. Kasrayan, Alex Johansson, Patrik Bergfors, Terese Sandström, Anders G. Bäckvall, Jan-Erling Mowbray, Sherry L. X-ray structure of Candida antarctica lipase A shows a novel lid structure and a likely mode of interfacial activation |
topic_facet |
lipase interfacial activation hydrolase X-ray structure substrate specificity Biochemistry and Molecular Biology Biokemi och molekylärbiologi |
description |
In nature, lipases (EC 3.1.1.3) catalyze the hydrolysis of triglycerides to form glycerol and fatty acids. Under the appropriate conditions, the reaction is reversible, and so biotechnological applications commonly make use of their capacity for esterification as well as for hydrolysis of a wide variety of compounds. In the present paper, we report the X-ray structure of lipase A from Candida antarctica, solved by single isomorphous replacement with anomalous scattering, and refined to 2.2-A resolution. The structure is the first from a novel family of lipases. Contrary to previous predictions, the fold includes a well-defined lid as well as a classic alpha/beta hydrolase domain. The catalytic triad is identified as Ser184, Asp334 and His366, which follow the sequential order considered to be characteristic of lipases; the serine lies within a typical nucleophilic elbow. Computer docking studies, as well as comparisons to related structures, place the carboxylate group of a fatty acid product near the serine nucleophile, with the long lipid tail closely following the path through the lid that is marked by a fortuitously bound molecule of polyethylene glycol. For an ester substrate to bind in an equivalent fashion, loop movements near Phe431 will be required, suggesting the primary focus of the conformational changes required for interfacial activation. Such movements will provide virtually unlimited access to solvent for the alcohol moiety of an ester substrate. The structure thus provides a basis for understanding the enzyme's preference for acyl moieties with long, straight tails, and for its highly promiscuous acceptance of widely different alcohol and amine moieties. An unconventional oxyanion hole is observed in the present structure, although the situation may change during interfacial activation. |
format |
Article in Journal/Newspaper |
author |
Ericsson, Daniel J. Kasrayan, Alex Johansson, Patrik Bergfors, Terese Sandström, Anders G. Bäckvall, Jan-Erling Mowbray, Sherry L. |
author_facet |
Ericsson, Daniel J. Kasrayan, Alex Johansson, Patrik Bergfors, Terese Sandström, Anders G. Bäckvall, Jan-Erling Mowbray, Sherry L. |
author_sort |
Ericsson, Daniel J. |
title |
X-ray structure of Candida antarctica lipase A shows a novel lid structure and a likely mode of interfacial activation |
title_short |
X-ray structure of Candida antarctica lipase A shows a novel lid structure and a likely mode of interfacial activation |
title_full |
X-ray structure of Candida antarctica lipase A shows a novel lid structure and a likely mode of interfacial activation |
title_fullStr |
X-ray structure of Candida antarctica lipase A shows a novel lid structure and a likely mode of interfacial activation |
title_full_unstemmed |
X-ray structure of Candida antarctica lipase A shows a novel lid structure and a likely mode of interfacial activation |
title_sort |
x-ray structure of candida antarctica lipase a shows a novel lid structure and a likely mode of interfacial activation |
publisher |
Uppsala universitet, Strukturell molekylärbiologi |
publishDate |
2008 |
url |
http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-14221 https://doi.org/10.1016/j.jmb.2007.10.079 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_relation |
Journal of Molecular Biology, 0022-2836, 2008, 376:1, s. 109-119 http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-14221 doi:10.1016/j.jmb.2007.10.079 PMID 18155238 ISI:000253181500011 |
op_rights |
info:eu-repo/semantics/openAccess |
op_doi |
https://doi.org/10.1016/j.jmb.2007.10.079 |
container_title |
Journal of Molecular Biology |
container_volume |
376 |
container_issue |
1 |
container_start_page |
109 |
op_container_end_page |
119 |
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1766273964220350464 |