Kinetic and inhibition studies of cinnamoyl-CoA reductase 1 from Arabidopsis thaliana
| Autor: | Jacqueline Grima-Pettenati, P. Saint-Aguet, Michel Baltas, Florence Bedos-Belval, L. Roussel, C. Lapeyre, M. Maturano, Hubert Duran |
|---|---|
| Přispěvatelé: | Synthèse et Physico-Chimie de Molécules d'Intérêt Biologique (SPCMIB), Institut de Chimie de Toulouse (ICT-FR 2599), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut de mécanique des fluides de Toulouse (IMFT), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées, inconnu, Inconnu |
| Rok vydání: | 2005 |
| Předmět: |
0106 biological sciences
Physiology Coenzyme A Arabidopsis Organophosphonates Plant Science Biology Reductase Thioester 01 natural sciences Substrate Specificity 03 medical and health sciences chemistry.chemical_compound Biosynthesis Genetics [CHIM]Chemical Sciences ComputingMilieux_MISCELLANEOUS 030304 developmental biology chemistry.chemical_classification 0303 health sciences Molecular Structure Substrate (chemistry) Aldehyde Oxidoreductases Enzyme assay Recombinant Proteins Kinetics Enzyme chemistry Biochemistry biology.protein Cinnamoyl-CoA reductase Acyl Coenzyme A 010606 plant biology & botany |
| Zdroj: | Plant Physiology and Biochemistry Plant Physiology and Biochemistry, Elsevier, 2005, 43 (8), pp.746-753. ⟨10.1016/j.plaphy.2005.06.003⟩ |
| ISSN: | 0981-9428 |
| DOI: | 10.1016/j.plaphy.2005.06.003⟩ |
| Popis: | Cinnamoyl coenzyme A reductase (CCR, EC 1.2.1.44), one of the key enzymes in the biosynthesis of lignin monomers, catalyzes the NADPH-dependent reduction of cinnamoyl-CoA esters to their corresponding cinnamaldehydes. AtCCR1, one of the two distinct isoforms isolated from Arabidopsis thaliana, was shown to be involved in lignin biosynthesis during development. Here, we report on the purification of the recombinant AtCCR1 protein expressed in Escherichia coli and the subsequent determination of its kinetic properties (K(m) and k(cat)/K(m) values) towards its main substrates i.e. feruloyl-CoA, sinapoyl-CoA, and p-coumaroyl-CoA esters. In addition, the potential inhibitory effect of five substrate-like analogs possessing an N-acetylcysteamine thioester group was tested on CCR activity using either feruloyl-CoA or sinapoyl-CoA as substrates. The K(i) values were in the range of 4.4-502 microM and the type of inhibition was found to be either uncompetitive or noncompetitive. Interestingly, for compounds 3 and 5, the type of inhibition was found to be different depending on the substrate used to monitor the enzyme activity. The best inhibitors were those possessing the feruloyl (compound 3) and sinapoyl (compound 5) aromatic moiety (4.1 and 7.1 microM) while the enzyme activity was monitored using the corresponding substrates. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect