5BSR

Crystal structure of 4-coumarate:CoA ligase complexed with adenosine monophosphate and Coenzyme A

Summary for 5BSR

• Entry DOI: 10.2210/pdb5bsr/pdb
• Related: 5BSM 5BST 5BSU 5BSV 5BSW
• Descriptor: 4-coumarate--CoA ligase 2, COENZYME A, ADENOSINE MONOPHOSPHATE, ... (5 entities in total)
• Functional Keywords: 4-coumarate:coa ligase, ligase
• Biological source: Nicotiana tabacum (Common tobacco)
• Total number of polymer chains: 1
• Total formula weight: 61028.78

Authors

Li, Z.,Nair, S.K. (deposition date: 2015-06-02, release date: 2016-05-11, Last modification date: 2024-03-06)

Primary citation

Li, Z.,Nair, S.K.
Structural Basis for Specificity and Flexibility in a Plant 4-Coumarate:CoA Ligase.
Structure, 23:2032-2042, 2015

PubMed Abstract: Plant 4-coumarate:CoA ligase (4CL) serves as a central catalyst in the phenylpropanoid pathway that provides precursors for numerous metabolites and regulates carbon flow. Here, we present several high-resolution crystal structures of Nicotiana tabacum 4CL isoform 2 (Nt4CL2) in complex with Mg(2+) and ATP, with AMP and coenzyme A (CoA), and with three different hydroxycinnamate-AMP intermediates: 4-coumaroyl-AMP, caffeoyl-AMP, and feruloyl-AMP. The Nt4CL2-Mg(2+)-ATP structure is captured in the adenylate-forming conformation, whereas the other structures are in the thioester-forming conformation. These structures represent a rare example of an ANL enzyme visualized in both conformations, and also reveal the binding determinants for both CoA and the hydroxycinnamate substrate. Kinetic studies of structure-based variants were used to identify residues crucial to catalysis, ATP binding, and hydroxycinnamate specificity. Lastly, we characterize a deletion mutant of Nt4CL2 that possesses the unusual sinapinate-utilizing activity. These studies establish a molecular framework for the engineering of this versatile biocatalyst.

PubMed: 26412334
DOI: 10.1016/j.str.2015.08.012

Experimental method

X-RAY DIFFRACTION (1.5 Å)