Probing the Interactions of Early Polyketide Intermediates with the Actinorhodin ACP from S. coelicolor A3(2)

Evans, Simon; Williams, Christopher; Arthur, Christopher J.; Płoskoń, Eliza; Wattana-Amorn, Pakorn; Cox, Russell J.; Crosby, John; Willis, Christine L.; Simpson, Thomas J.; Crump, Matthew P.

doi:10.1016/j.jmb.2009.03.072

Cited by 51 publications

(114 citation statements)

References 60 publications

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“…Analogous carrier proteins from polyketide synthetases can sequester their molecular cargo, although the biochemical relevance of this mechanism is not known 31, 32, 33. In the case of PCPs, the Ppant arm does not appear to interact appreciably with the protein core nor to alter the tertiary structure of the PCP in a significant way 21, 34.…”

Section: The Peptidyl Carrier Proteinmentioning

confidence: 99%

New Structural Data Reveal the Motion of Carrier Proteins in Nonribosomal Peptide Synthesis

et al. 2016

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The nonribosomal peptide synthetases (NRPSs) are one of the most promising resources for the production of new bioactive molecules. The mechanism of NRPS catalysis is based around sequential catalytic domains: these are organized into modules, where each module selects, modifies, and incorporates an amino acid into the growing peptide. The intermediates formed during NRPS catalysis are delivered between enzyme centers by peptidyl carrier protein (PCP) domains, which makes PCP interactions and movements crucial to NRPS mechanism. PCP movement has been linked to the domain alternation cycle of adenylation (A) domains, and recent complete NRPS module structures provide support for this hypothesis. However, it appears as though the A domain alternation alone is insufficient to account for the complete NRPS catalytic cycle and that the loaded state of the PCP must also play a role in choreographing catalysis in these complex and fascinating molecular machines.

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Section: The Peptidyl Carrier Proteinmentioning

confidence: 99%

New Structural Data Reveal the Motion of Carrier Proteins in Nonribosomal Peptide Synthesis

et al. 2016

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“…These findings are also in agreement with recent studies on ACP6 of DEBS 25 and the actinorhodin ACP. 26,27 Influence of the growing polyketide chain on protein-protein interactions during intermodular chain translocation…”

Section: Preparation and Structural Analysis Of Holo- Trifluoromethymentioning

confidence: 99%

Probing the interactions of an acyl carrier protein domain from the 6‐deoxyerythronolide B synthase

et al. 2011

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The assembly-line architecture of polyketide synthases (PKSs) provides an opportunity to rationally reprogram polyketide biosynthetic pathways to produce novel antibiotics. A fundamental challenge toward this goal is to identify the factors that control the unidirectional channeling of reactive biosynthetic intermediates through these enzymatic assembly lines. Within the catalytic cycle of every PKS module, the acyl carrier protein (ACP) first collaborates with the ketosynthase (KS) domain of the paired subunit in its own homodimeric module so as to elongate the growing polyketide chain and then with the KS domain of the next module to translocate the newly elongated polyketide chain. Using NMR spectroscopy, we investigated the features of a structurally characterized ACP domain of the 6-deoxyerythronolide B synthase that contribute to its association with its KS translocation partner. Not only were we able to visualize selective protein-protein interactions between the two partners, but also we detected a significant influence of the acyl chain substrate on this interaction. A novel reagent, CF 3 -S-ACP, was developed as a 19 F NMR spectroscopic probe of protein-protein interactions. The implications of our findings for understanding intermodular chain translocation are discussed.

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“…Acyl carrier protein (ACP) labeling with 4’-phosphopantetheine (ppant), conjugated to a tag of choice by its transferase (PPTase) represents one of the most flexible covalent protein labeling methods, as illustrated by the application of ACP tagged peptides 7 , for bio-gel formation 8 , and ACP-dependent protein immobilization 9 . Labeling ACP and ACP fusion proteins with ppant analogs is also successfully leveraged for visualization 10,11 , isolation 12 , functional 13 , and structural 14,15 studies of carrier protein-dependent biosynthetic enzymes 16 . Yet further advancement of these tools is hampered by an inability to easily reverse ppant attachment.…”

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confidence: 99%

“…Biosynthetic intermediates with varied chemical structures participate in intermolecular interactions with ACP that modulate substrate dynamics and ACP structure 9,15 . The nature of these ACP-substrate interactions depends on the chemical structure of the biosynthetic intermediate and can vary with respect to chain length and oxidation state 23 .…”

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confidence: 99%

Reversible labeling of native and fusion-protein motifs

et al. 2012

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The reversible covalent attachment of chemical probes to proteins has long been sought as a means to visualize and manipulate proteins. Here we demonstrate the full reversibility of post-translational custom pantetheine modification of E. coli acyl carrier protein (ACP) for visualization and functional studies. We utilize this iterative enzymatic methodology in vitro for reversible labeling variants and apply these tools to Nuclear Magnetic Resonance (NMR) structural studies of protein-substrate interactions.

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Probing the Interactions of Early Polyketide Intermediates with the Actinorhodin ACP from S. coelicolor A3(2)

Cited by 51 publications

References 60 publications

New Structural Data Reveal the Motion of Carrier Proteins in Nonribosomal Peptide Synthesis

New Structural Data Reveal the Motion of Carrier Proteins in Nonribosomal Peptide Synthesis

Probing the interactions of an acyl carrier protein domain from the 6‐deoxyerythronolide B synthase

Reversible labeling of native and fusion-protein motifs

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