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The substrate specificity-determining amino acid code of 4-coumarate:CoA ligase
Open accessAbstract
To reveal the structural principles determining substrate specificity of 4-coumarate:CoA ligase (4CL), the crystal structure of the phenylalanine activation domain of gramicidin S synthetase was used as a template for homology modeling. According to our model, 12 amino acid residues lining the Arabidopsis 4CL isoform 2 (At4CL2) substrate binding pocket (SBP) function as a signature motif generally determining 4CL substrate specificity. We used this substrate specificity code to create At4CL2 gain-of-function mutants. By increasing the space within the SBP we generated ferulic- and sinapic acid-activating At4CL2 variants. Increasing the hydrophobicity of the SBP resulted in At4CL2 variants with strongly enhanced conversion of cinnamic acid. These enzyme variants are suitable tools for investigating and influencing metabolic channeling mediated by 4CL. Knowledge of the 4CL specificity code will facilitate the prediction of substrate preference of numerous, still uncharacterized 4CL-like proteins.
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Cited by
The Phenylpropanoid Pathway in Arabidopsis
Fraser, C. M., & Chapple, C. (2011) · The Arabidopsis Book · doi:10.1199/tab.0152 · Cited by 860
Chen, H.-C., Song, J., Wang, J. P., Lin, Y.-C., Ducoste, J., Shuford, C. M., Liu, J., Li, Q., Shi, R., Nepomuceno, A., et al. (2014) · The Plant Cell · doi:10.1105/tpc.113.119685 · Cited by 80
Early phenylpropanoid biosynthetic steps in Cannabis sativa: link between genes and metabolites.
Docimo, T., Consonni, R., Coraggio, I., & Mattana, M. (2013) · International Journal of Molecular Sciences · doi:10.3390/ijms140713626 · Cited by 39
Metadata
- Authors
- Schneider, K., Hövel, K., Witzel, K., Hamberger, B., Schomburg, D., Kombrink, E., & Stuible, H.-P.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 2003-06-20
- Volume
- 100
- Issue
- 14
- Pages
- 8601-8606
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W2058442404
- Cited by
- 164
- References
- 0
- FWCI
- 1.82
- Type
- journal article
- Language
- en
- Open access
- green
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- Schneider, K., Hövel, K., Witzel, K., Hamberger, B., Schomburg, D., Kombrink, E., & Stuible, H.-P. (2003). The substrate specificity-determining amino acid code of 4-coumarate:CoA ligase. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.1430550100
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