A new buckwheat dihydroflavonol 4-reductase (DFR), with a unique substrate binding structure, has altered substrate specificity
| العنوان: | A new buckwheat dihydroflavonol 4-reductase (DFR), with a unique substrate binding structure, has altered substrate specificity |
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| المؤلفون: | Wataru Tsuchiya, Kenjiro Katsu, Kiyoshi Furukawa, Yasuo Yasui, Toshimasa Yamazaki, Seiji Kubota, Toshiyuki Komori, Amanda R. Walker, Tomomi Hisano, Rintaro Suzuki, Noritoshi Inagaki, Katsuhiro Matsui, Motoyuki Koshio |
| المصدر: | SC10201804160188 NARO成果DBa This is an open-access article distributed. CC BY4.0 BMC Plant Biology, Vol 17, Iss 1, Pp 1-14 (2017) BMC Plant Biology |
| بيانات النشر: | Springer Science and Business Media LLC, 2017. |
| سنة النشر: | 2017 |
| مصطلحات موضوعية: | 0106 biological sciences, 0301 basic medicine, Recombinant protein, Population, Sequence alignment, Plant Science, Biology, 01 natural sciences, Substrate Specificity, Anthocyanins, 03 medical and health sciences, lcsh:Botany, Complementary DNA, Proanthocyanidins, Amino Acid Sequence, Asparagine, Fagopyrum esculentum, education, Peptide sequence, Phylogeny, Plant Proteins, chemistry.chemical_classification, education.field_of_study, Substrate (chemistry), biology.organism_classification, lcsh:QK1-989, Amino acid, 3D structure modelling, Alcohol Oxidoreductases, 030104 developmental biology, chemistry, Biochemistry, Flavonoid, Substrate preference, Sequence Alignment, Fagopyrum, Research Article, 010606 plant biology & botany |
| الوصف: | Background Dihydroflavonol 4-reductase (DFR) is the key enzyme committed to anthocyanin and proanthocyanidin biosynthesis in the flavonoid biosynthetic pathway. DFR proteins can catalyse mainly the three substrates (dihydrokaempferol, dihydroquercetin, and dihydromyricetin), and show different substrate preferences. Although relationships between the substrate preference and amino acids in the region responsible for substrate specificity have been investigated in several plant species, the molecular basis of the substrate preference of DFR is not yet fully understood. Results By using degenerate primers in a PCR, we isolated two cDNA clones that encoded DFR in buckwheat (Fagopyrum esculentum). Based on sequence similarity, one cDNA clone (FeDFR1a) was identical to the FeDFR in DNA databases (DDBJ/Gen Bank/EMBL). The other cDNA clone, FeDFR2, had a similar sequence to FeDFR1a, but a different exon-intron structure. Linkage analysis in an F2 segregating population showed that the two loci were linked. Unlike common DFR proteins in other plant species, FeDFR2 contained a valine instead of the typical asparagine at the third position and an extra glycine between sites 6 and 7 in the region that determines substrate specificity, and showed less activity against dihydrokaempferol than did FeDFR1a with an asparagine at the third position. Our 3D model suggested that the third residue and its neighbouring residues contribute to substrate specificity. FeDFR1a was expressed in all organs that we investigated, whereas FeDFR2 was preferentially expressed in roots and seeds. Conclusions We isolated two buckwheat cDNA clones of DFR genes. FeDFR2 has unique structural and functional features that differ from those of previously reported DFRs in other plants. The 3D model suggested that not only the amino acid at the third position but also its neighbouring residues that are involved in the formation of the substrate-binding pocket play important roles in determining substrate preferences. The unique characteristics of FeDFR2 would provide a useful tool for future studies on the substrate specificity and organ-specific expression of DFRs. Electronic supplementary material The online version of this article (10.1186/s12870-017-1200-6) contains supplementary material, which is available to authorized users. |
| تدمد: | 1471-2229 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::5d6e76e5db894307ac4bf2decac312c9Test https://doi.org/10.1186/s12870-017-1200-6Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....5d6e76e5db894307ac4bf2decac312c9 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 14712229 |
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