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Shifting the Hydrolysis Equilibrium of Substrate Loaded Acyl Carrier Proteins
- Source :
- Biochemistry, vol 58, iss 34, Biochemistry
- Publication Year :
- 2019
- Publisher :
- American Chemical Society (ACS), 2019.
-
Abstract
- Acyl carrier proteins (ACP)s transport intermediates through many primary and secondary metabolic pathways. Studying the effect of substrate identity on ACP structure has been hindered by the lability of the thioester bond that attaches acyl substrates to the 4’-phosphopantetheine cofactor of ACP. Here we show that an acyl acyl-carrier protein synthetase (AasS) can be used in real time to shift the hydrolysis equilibrium towards favoring acyl-ACP during solution NMR spectroscopy. Only 0.005 molar equivalents of AasS enables one week of stability to palmitoyl-AcpP from Escherichia coli. 2D NMR spectra enabled with this method revealed that the tethered palmitic acid perturbs nearly every secondary structural region of AcpP. This technique will allow previously unachievable structural studies of unstable acyl-ACP species, contributing to the understanding of these complex biosynthetic pathways.
- Subjects :
- Biochemistry & Molecular Biology
Protein Conformation
Stereochemistry
Medical Biochemistry and Metabolomics
Thioester
Biochemistry
Article
Cofactor
Medicinal and Biomolecular Chemistry
03 medical and health sciences
Hydrolysis
chemistry.chemical_compound
Protein structure
Escherichia coli
Acyl Carrier Protein
Genetics
chemistry.chemical_classification
0303 health sciences
biology
030302 biochemistry & molecular biology
Substrate (chemistry)
Nuclear magnetic resonance spectroscopy
chemistry
Pantetheine
biology.protein
lipids (amino acids, peptides, and proteins)
Biochemistry and Cell Biology
Phosphopantetheine
Two-dimensional nuclear magnetic resonance spectroscopy
Biotechnology
Subjects
Details
- ISSN :
- 15204995 and 00062960
- Volume :
- 58
- Database :
- OpenAIRE
- Journal :
- Biochemistry
- Accession number :
- edsair.doi.dedup.....23dba6eadd1023bab8d8cf4942e5c98b