Acyl Carrier Protein Cyanylation Delivers a Ketoacyl Synthase–Carrier Protein Cross-Link

Acyl carrier proteins (ACPs) are central hubs in polyketide and fatty acid biosynthetic pathways, but the fast motions of the ACP’s phosphopantetheine (Ppant) arm make its conformational dynamics difficult to capture using traditional spectroscopic approaches. Here we report that converting the term...

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Bibliographic Details
Published in:Biochemistry (Easton) 2017-05, Vol.56 (20), p.2533-2536
Main Authors: Thiele, Grace A. R, Friedman, Connie P, Tsai, Kathleen J. S, Beld, Joris, Londergan, Casey H, Charkoudian, Louise K
Format: Article
Language:English
Subjects:
active sites
acyl carrier protein
Acyl Carrier Protein - chemistry
biocatalysis
biochemical pathways
Chromatography, Gel
cyanides
Cyanides - chemistry
cysteine
Escherichia coli
Escherichia coli Proteins - chemistry
fatty acids
infrared spectroscopy
Polyketide Synthases - chemistry
polyketides
thiocyanates
thiols
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Summary:Acyl carrier proteins (ACPs) are central hubs in polyketide and fatty acid biosynthetic pathways, but the fast motions of the ACP’s phosphopantetheine (Ppant) arm make its conformational dynamics difficult to capture using traditional spectroscopic approaches. Here we report that converting the terminal thiol of Escherichia coli ACP’s Ppant arm into a thiocyanate activates this site to form a selective cross-link with the active site cysteine of its partner ketoacyl synthase (FabF). The reaction releases a cyanide anion, which can be detected by infrared spectroscopy. This represents a practical and generalizable method for obtaining and visualizing ACP–protein complexes relevant to biocatalysis and will be valuable in future structural and engineering studies.
ISSN:0006-2960
1520-4995
1520-4995
DOI:10.1021/acs.biochem.7b00219