Characterization of the cinnamoyl-CoA reductase (CCR) gene family in Populus tomentosa reveals the enzymatic active sites and evolution of CCR
- PMID: 27580618
- DOI: 10.1007/s00425-016-2591-6
Characterization of the cinnamoyl-CoA reductase (CCR) gene family in Populus tomentosa reveals the enzymatic active sites and evolution of CCR
Abstract
Two distinct cinnamoyl-coenzyme A reductases (CCRs) from Populus tomentosa were cloned and studied and active sites in CCRs were further identified based on sequence divergence, molecular simulation, and site-directed mutants. Cinnamoyl-coenzyme A (CoA) reductase (CCR) is the first committed gene in the lignin-specific pathway and plays a role in the lignin biosynthesis pathway. In this study, we cloned 11 genes encoding CCR or CCR-like proteins in Populus tomentosa. An enzymatic assay of the purified recombinant P. tomentosa (Pto) CCR and PtoCCR-like proteins indicated that only PtoCCR1 and PtoCCR7 had detectable activities toward hydroxycinnamoyl-CoA esters. PtoCCR1 exhibited specificity for feruloyl-CoA, with no detectable activity for any other hydroxycinnamoyl-CoA esters. However, PtoCCR7 catalyzed p-coumaroyl-CoA, caffeoyl-CoA, feruloyl-CoA, and sinapoyl-CoA with a preference for feruloyl-CoA. Site-directed mutations of selected amino acids divergent between PtoCCR1 and 7, combined with modeling and docking, showed that A132 in CCR7 combined with the catalytic triad might comprise the catalytic center. In CCR7, L192, F155, and H208 were identified as the substrate-binding sites, and site-directed mutations of these amino acids showed obvious changes in catalytic efficiency with respect to both feruloyl-CoA and sinapoyl-CoA. Mutant F155Y exhibited greater catalytic efficiency for sinapoyl-CoA compared with that of wild-type PtoCCR7. Finally, recent genome duplication events provided the foundation for CCR divergence. This study further identified the active sites in CCRs and the evolutionary process of CCRs in terrestrial plants.
Keywords: Binding sites; Cinnamoyl-coenzyme A reductase; Gene family; Populus tomentosa; Site-directed mutations.
Similar articles
-
Comparative and phylogenomic analyses of cinnamoyl-CoA reductase and cinnamoyl-CoA-reductase-like gene family in land plants.Plant Sci. 2011 Sep;181(3):249-57. doi: 10.1016/j.plantsci.2011.05.012. Epub 2011 May 26. Plant Sci. 2011. PMID: 21763535
-
Two distinct cinnamoyl-CoA reductases in Selaginella moellendorffii offer insight into the divergence of CCRs in plants.Planta. 2017 Jul;246(1):33-43. doi: 10.1007/s00425-017-2678-8. Epub 2017 Mar 20. Planta. 2017. PMID: 28321576
-
Clarification of cinnamoyl co-enzyme A reductase catalysis in monolignol biosynthesis of Aspen.Plant Cell Physiol. 2005 Jul;46(7):1073-82. doi: 10.1093/pcp/pci120. Epub 2005 May 3. Plant Cell Physiol. 2005. PMID: 15870094
-
Evolution of new enzymes by gene duplication and divergence.FEBS J. 2020 Apr;287(7):1262-1283. doi: 10.1111/febs.15299. FEBS J. 2020. PMID: 32250558 Free PMC article. Review.
-
On the natural diversity of phenylacylated-flavonoid and their in planta function under conditions of stress.Phytochem Rev. 2018;17(2):279-290. doi: 10.1007/s11101-017-9531-3. Epub 2017 Sep 8. Phytochem Rev. 2018. PMID: 29755304 Free PMC article. Review.
Cited by
-
Association of gene expression with syringyl to guaiacyl ratio in sugarcane lignin.Plant Mol Biol. 2021 May;106(1-2):173-192. doi: 10.1007/s11103-021-01136-w. Epub 2021 Mar 18. Plant Mol Biol. 2021. PMID: 33738678
-
De novo transcriptome and phytochemical analyses reveal differentially expressed genes and characteristic secondary metabolites in the original oolong tea (Camellia sinensis) cultivar 'Tieguanyin' compared with cultivar 'Benshan'.BMC Genomics. 2019 Apr 3;20(1):265. doi: 10.1186/s12864-019-5643-z. BMC Genomics. 2019. PMID: 30943892 Free PMC article.
-
A peroxisomal β-oxidative pathway contributes to the formation of C6-C1 aromatic volatiles in poplar.Plant Physiol. 2021 Jun 11;186(2):891-909. doi: 10.1093/plphys/kiab111. Plant Physiol. 2021. PMID: 33723573 Free PMC article.
-
Genome-wide identification, characterization, and genetic diversity of CCR gene family in Dalbergia odorifera.Front Plant Sci. 2022 Dec 19;13:1064262. doi: 10.3389/fpls.2022.1064262. eCollection 2022. Front Plant Sci. 2022. PMID: 36600926 Free PMC article.
-
Comparative Genomic Analysis of SAUR Gene Family, Cloning and Functional Characterization of Two Genes (PbrSAUR13 and PbrSAUR52) in Pyrus bretschneideri.Int J Mol Sci. 2022 Jun 24;23(13):7054. doi: 10.3390/ijms23137054. Int J Mol Sci. 2022. PMID: 35806062 Free PMC article.
References
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
