1. Structural investigation of the docking domain assembly from trans-AT polyketide synthases.
- Author
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Son SY, Bae DW, Kim E, Jeong BG, Kim MY, Youn SY, Yi S, Kim G, Hahn JS, Lee NK, Yoon YJ, and Cha SS
- Subjects
- Crystallography, X-Ray, Bacterial Proteins chemistry, Bacterial Proteins metabolism, Bacterial Proteins genetics, Models, Molecular, Protein Domains, beta-Lactamases chemistry, beta-Lactamases metabolism, beta-Lactamases genetics, Protein Binding, Polyketide Synthases chemistry, Polyketide Synthases metabolism, Polyketide Synthases genetics, Acyltransferases metabolism, Acyltransferases chemistry, Acyltransferases genetics
- Abstract
Docking domains (DDs) located at the C- and N-termini of polypeptides play a crucial role in directing the assembly of polyketide synthases (PKSs), which are multienzyme complexes. Here, we determined the crystal structure of a complex comprising the C-terminal DD (
C DDMlnB ) and N-terminal DD (N DDMlnC ) of macrolactin trans-acyltransferase (AT) PKS that were fused to a functional enzyme, AmpC EC2 β-lactamase. Interface analyses of theC DDMlnB /N DDMlnC complex revealed the molecular intricacies in the core section underpinning the precise DD assembly. Additionally, circular dichroism and steady-state kinetics demonstrated that the formation of theC DDMlnB /N DDMlnC complex had no influence on the structural and functional fidelity of the fusion partner, AmpC EC2. This inspired us to apply theC DDMlnB /N DDMlnC assembly to metabolon engineering. Indeed, DD assembly induced the formation of a complex between 4-coumarate-CoA ligase and chalcone synthase both involved in flavonoid biosynthesis, leading to a remarkable increase in naringenin production in vitro., Competing Interests: Declaration of interests D.-W.B., E.K., B.-G.J., S.-J.Y., N.K.L., J.-S.H., S.-S.C., and Y.J.Y. have filed a patent for employing docking domains to assemble an artificial metabolon, as described in this study., (Copyright © 2024 Elsevier Inc. All rights reserved.)- Published
- 2024
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