Work

A Nuclear Magnetic Resonance Method for Probing Molecular Influences of Substrate Loading in Nonribosomal Peptide Synthetase Carrier Proteins

Year: 2015

Type: article

Abstract: Carrier proteins (CPs) play a central role in nonribosomal peptide synthetases (NRPSs) as they shuttle covalently attached substrates between active sites. Understanding how the covalent attachment of a substrate (loading) influences the molecular properties of CPs is key to determining the mechanism of NRPS synthesis. However, structural studies have been impaired by substrate hydrolysis. Here, we used nuclear magnetic resonance spectroscopy to monitor substrate loading of a CP and to overcome hydrolysis. Our results reveal the spectroscopic signature of substrate loading and provide evidence of molecular communication between an NRPS carrier protein and its covalently attached substrate.

Source: Biochemistry

DOI: 10.1021/bi501433r

Authors: Andrew C. Goodrich, Dominique P. Frueh

Institutions: Johns Hopkins University, Johns Hopkins Medicine

Language: English

Topic: Microbial Natural Products and Biosynthesis

Subfield: Pharmacology

Field: Medicine

Domain: Health Sciences

Open Access status: green

Funders: American Heart Association, National Institute of General Medical Sciences, National Center for Research Resources

Awards: G151248532, Biochemistry, Cellular and Molecular Biology Program, Purchase of a 600 MHz NMR Console and Probes, G691550182