Employing 25-Residue Docking Motifs from Modular Polyketide Synthases as Orthogonal Protein Connectors

Jessica L. Meinke, Anna J. Simon, Drew T. Wagner, Barrett R. Morrow, Shaochen You, Andrew D. Ellington, Adrian T. Keatinge-Clay

Research output: Contribution to journalArticle

Abstract

The proteins of trans-acyltransferase modular polyketide synthases (PKSs) self-organize into assembly lines, enabling the multienzyme biosynthesis of complex organic molecules. Docking domains comprised of ∼25 residues at the C- and N-termini of these polypeptides (CDDs and NDDs) help drive this association through the formation of four-helix bundles. Molecular connectors like these are desired in synthetic contexts, such as artificial biocatalytic systems and biomaterials, to orthogonally join proteins. Here, the ability of six CDD/NDD pairs to link non-PKS proteins is examined using green fluorescent protein (GFP) variants. As observed through size-exclusion chromatography and Förster resonance energy transfer (FRET), matched but not mismatched pairs of Venus+CDD and NDD+mTurquoise2 fusion proteins associate with low micromolar affinities.

Original languageEnglish (US)
Pages (from-to)2017-2024
Number of pages8
JournalACS Synthetic Biology
Volume8
Issue number9
DOIs
StatePublished - Sep 20 2019

Fingerprint

Polyketide Synthases
Proteins
Venus
Multienzyme Complexes
Energy Transfer
Biocompatible Materials
Green Fluorescent Proteins
Acyltransferases
Gel Chromatography
Size exclusion chromatography
Biosynthesis
Energy transfer
Polypeptides
Peptides
Fusion reactions
Biomaterials
Association reactions
Molecules

Keywords

  • FRET
  • docking domain
  • fluorescent proteins
  • four-helix bundle
  • orthogonal connectors
  • polyketide synthase

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

Cite this

Employing 25-Residue Docking Motifs from Modular Polyketide Synthases as Orthogonal Protein Connectors. / Meinke, Jessica L.; Simon, Anna J.; Wagner, Drew T.; Morrow, Barrett R.; You, Shaochen; Ellington, Andrew D.; Keatinge-Clay, Adrian T.

In: ACS Synthetic Biology, Vol. 8, No. 9, 20.09.2019, p. 2017-2024.

Research output: Contribution to journalArticle

Meinke, Jessica L. ; Simon, Anna J. ; Wagner, Drew T. ; Morrow, Barrett R. ; You, Shaochen ; Ellington, Andrew D. ; Keatinge-Clay, Adrian T. / Employing 25-Residue Docking Motifs from Modular Polyketide Synthases as Orthogonal Protein Connectors. In: ACS Synthetic Biology. 2019 ; Vol. 8, No. 9. pp. 2017-2024.
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