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Expression of 5,8-LDS of Aspergillus fumigatus and its dioxygenase domain : a comparison with 7,8-LDS, 10-dioxygenase, and cyclooxygenase
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- Hoffmann, Inga, 1984- (author)
- Uppsala universitet,Institutionen för farmaceutisk biovetenskap,Biokemisk farmakologi
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- Jernerén, Fredrik (author)
- Uppsala universitet,Institutionen för farmaceutisk biovetenskap,Biokemisk farmakologi
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- Garscha, Ulrike (author)
- Uppsala universitet,Institutionen för farmaceutisk biovetenskap,Biokemisk farmakologi
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- (creator_code:org_t)
- Elsevier BV, 2011
- 2011
- English.
- In: Archives of Biochemistry and Biophysics. - : Elsevier BV. - 0003-9861 .- 1096-0384. ; 506:2, s. 216-222
- Journal article (peer-reviewed)
Abstract
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- 5,8-Linoleate diol synthase (5,8-LDS) of Aspergillus fumigatus was cloned, expressed, and compared with 7,8-LDS of the Take-all fungus. Replacements of Tyr and Cys in the conserved YRWH and FXXGPHXCLG sequences abolished 8R-dioxygenase (8-DOX) and hydroperoxide isomerase activities, respectively. The predicted α-helices of LDS were aligned with α-helices of cyclooxygenase-1 (COX-1) to identify the 8-DOX domains. N-terminal expression constructs of 5,8- and 7,8-LDS (674 of 1079, and 673 of 1165 residues), containing one additional α-helix compared to cyclooxygenase-1, yielded prominent 8R-DOX activities with apparently unchanged or slightly lower substrate affinities, respectively. Val-328 of 5,8-LDS did not influence the position of oxygenation in contrast to the homologous residues Val-349 of COX-1 and Leu-384 of 10R-dioxygenase. We conclude that ∼675 amino acids are sufficient to support 8-DOX activity.
Subject headings
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi -- Medicinsk bioteknologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology -- Medical Biotechnology (hsv//eng)
Keyword
- Cytochrome P450
- fusion protein
- heme-dependent peroxidase
- hydroperoxide isomerase
- LC-MS/MS
- oxylipins
- Biochemistry
- Biokemi
- Biokemisk farmakologi
- Biochemical Pharmacology
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- ref (subject category)
- art (subject category)
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