Repository logo
 
Publication

Isolation and characterization of a cDNA clone from Catharanthus roseus encoding NADPH: cytochrome P-450 reductase, an enzyme essential for reactions catalysed by cytochrome P-450 mono-oxygenases in plants

dc.contributor.authorMeijer, Annemarie H.
dc.contributor.authorCardoso, Inês Lopes
dc.contributor.authorVoskuilen, John Th.
dc.contributor.authorde Waal, Anthony
dc.contributor.authorVerpoorte, Robert
dc.contributor.authorHoge, J. Harry C.
dc.date.accessioned2020-03-05T18:12:42Z
dc.date.available2020-03-05T18:12:42Z
dc.date.issued1993
dc.date.updated2020-02-28T17:07:56Z
dc.description.abstractThe membrane‐bound flavoprotein NADPH:cytochrome P‐450 (cytochrome c) reductase, that functions in electron transfer to cytochrome P‐450 mono‐oxygenases, was purified from a cell suspension culture of the higher plant Catheranthus roseus. Anti‐serum raised against the purified protein was found to inhibit NADPH:cytochrome c reductase activity as well as the activities of the cytochrome P‐450 enzymes geraniol 10‐hydroxylase and trans‐cinnamate 4‐hydroxylase, which are involved in alkaloid biosynthesis and phenylpropanoid biosynthesis, respectively. Immunoscreening of a C. roseus cDNA expression library resulted in the isolation of a partial NADPH: cytochrome P‐450 reductase cDNA clone, which was identified on the basis of sequence homology with NADPH:cytochrome P‐450 reductases from yeast and animal species. The identity of the cDNA was confirmed by expression in Escherichia coli as a functional protein capable of NADPH‐dependent reduction of cytochrome c and neotetrazolium, two in vitro substrates for the reductase. The N‐terminal sequence of the reductase, which was not present in the cDNA clone, was determined from a genomic NADPH: cytochrome P‐450 reductase clone. It was demonstrated that the reductase probably is encoded by a single copy gene. A sequence comparison of this plant NADPH:cytochrome P‐450 reductase with the corresponding enzymes from yeast and animal species showed that functional domains involved in binding of the cofactors FMN, FAD and NADPH are highly conserved between all kingdoms. In C. roseus cell cultures a rapid increase of the reductase steady state mRNA level was observed after the addition of fungal elicitor preparations that are known to induce cytochrome P‐450‐dependent biosynthetic pathways.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.slugcv-prod-368666
dc.identifier.urihttp://hdl.handle.net/10284/8619
dc.language.isoengpt_PT
dc.subjectNADPH:cytochrome P-450 reductasept_PT
dc.subjectCatharanthus roseuspt_PT
dc.subjectTerpenoid indole alkaloidspt_PT
dc.titleIsolation and characterization of a cDNA clone from Catharanthus roseus encoding NADPH: cytochrome P-450 reductase, an enzyme essential for reactions catalysed by cytochrome P-450 mono-oxygenases in plantspt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.conferencePlacePaíses Baixospt_PT
oaire.citation.endPage60pt_PT
oaire.citation.issue1pt_PT
oaire.citation.startPage47pt_PT
oaire.citation.titlePlant Journalpt_PT
oaire.citation.volume4pt_PT
person.familyNameLopes Cardoso
person.givenNameInês
person.identifier.ciencia-idF21F-16B4-3715
person.identifier.orcid0000-0002-0693-9831
person.identifier.ridM-7156-2013
person.identifier.scopus-author-id54973754300
rcaap.cv.cienciaidF21F-16B4-3715 | Maria Inês de Avelar Lopes Cardoso
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublicationf39fa18f-2d23-4690-bfa2-f8ba53e40775
relation.isAuthorOfPublication.latestForDiscoveryf39fa18f-2d23-4690-bfa2-f8ba53e40775

Files

Original bundle
Now showing 1 - 1 of 1
Loading...
Thumbnail Image
Name:
MeijerLopesCardoso93.pdf
Size:
2.14 MB
Format:
Adobe Portable Document Format
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
1.64 KB
Format:
Item-specific license agreed upon to submission
Description: