Analysis and engineering of substrate shuttling by the acyl carrier protein (ACP) in fatty acid synthases (FASs)
Autor: | Emanuele Rossini, Martin Grininger, Gerhard Hummer, Maja Klaus, Jan Gajewski |
---|---|
Rok vydání: | 2018 |
Předmět: |
0301 basic medicine
Fatty Acid Synthases Enzyme complex Saccharomyces cerevisiae Proteins Saccharomyces cerevisiae Protein Engineering Catalysis 03 medical and health sciences Materials Chemistry Acyl Carrier Protein Molecule Point Mutation Protein Interaction Domains and Motifs Amino Acid Sequence Binding site Peptide sequence chemistry.chemical_classification Binding Sites biology Fatty Acids Metals and Alloys Fatty acid General Chemistry Protein engineering Surfaces Coatings and Films Electronic Optical and Magnetic Materials Fatty Acid Synthase Type I Molecular Docking Simulation Acyl carrier protein Fatty acid synthase 030104 developmental biology Enzyme Biochemistry chemistry Covalent bond Ceramics and Composites biology.protein Thermodynamics Function (biology) |
Zdroj: | Chemical communications (Cambridge, England). 54(82) |
ISSN: | 1364-548X |
Popis: | In the large enzyme complexes of natural biosynthetic pathways, molecules are assembled like in a factory. Carrier domains shuttle substrates and intermediates as covalently attached cargo within the enzyme complex between active sites. The physical confinement of the reaction increases reaction rates and hinders pathway branching. Alternating interactions of substrate-loaded carrier domains with different catalytic domains modulate the chemical environment. In this study, we aim at assessing the impact of domain-domain interactions (DDIs) on the reaction progress of a multienzyme type I fatty acid synthase (FAS) in quantitative terms. We modulate DDIs by single interface mutations, and read out the impact on substrate shuttling by recording fatty acid (FA) chain length product spectra and FAS activities. Our data show that even single interface point mutations can severely affect FA synthesis. With molecular dynamics simulations and modeling, we relate the mutation effects to specific alterations in the molecular interaction networks and domain-domain binding energetics. Some of the presented mutations induce the synthesis of short-chain FAs. These compounds are important commodity products and potent precursors for microbial biofuel production. |
Databáze: | OpenAIRE |
Externí odkaz: |