Nonribosomal peptide synthetases serve as multidomain protein templates for producing a wealth of pharmaceutically important natural products. For the correct assembly of the desired natural product the interactions between the different catalytic centres and the reaction intermediates bound to the peptidyl carrier protein must be precisely controlled at spatial and temporal levels. We have investigated the interplay between the adenylation (A) domain and the peptidyl carrier protein in the gramicidin S synthetase I (EC 5.1.1.11) via partial tryptic digests, native PAGE and gel-filtration analysis, as well as by chemical labeling experiments. Our data imply that the 4'-phosphopantetheine moiety of the peptidyl carrier protein changes its position as a result of a conformational change in the A domain, which is induced by the binding of an amino acyl adenylate mimic. The productive interaction between the two domains at the stage of the amino acyl transfer onto the 4'-phosphopantetheine moiety is accompanied by a highly compact protein conformation of the holo-protein. These results provide the first biochemical evidence for the occurrence of conformational changes in the cross-talk between A and peptidyl carrier protein domains of a multidomain nonribosomal peptide synthetase.
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http://dx.doi.org/10.1111/j.1742-4658.2009.07551.x | DOI Listing |
Nature
December 2024
Department of Chemistry and Biochemistry, University of California San Diego, La Jolla, CA, USA.
Non-ribosomal peptide synthetases are assembly line biosynthetic pathways that are used to produce critical therapeutic drugs and are typically arranged as large multi-domain proteins called megasynthetases. They synthesize polypeptides using peptidyl carrier proteins that shuttle each amino acid through modular loading, modification and elongation steps, and remain challenging to structurally characterize, owing in part to the inherent dynamics of their multi-domain and multi-modular architectures. Here we have developed site-selective crosslinking probes to conformationally constrain and resolve the interactions between carrier proteins and their partner enzymatic domains.
View Article and Find Full Text PDFJ Food Drug Anal
September 2024
Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, UPM, 43400, Serdang, Selangor, Malaysia.
Int J Biol Macromol
January 2025
College of Food Science, Northeast Agricultural University, Harbin, Heilongjiang 150030, China; Research and Product Development Center, Shandong Guohong Biotechnology Company Limited, Liaocheng, Shandong 252899, China. Electronic address:
PLoS Negl Trop Dis
November 2024
Laboratório de Biologia das Interações, Instituto Oswaldo Cruz/Fiocruz, Rio de Janeiro, Rio de Janeiro, Brazil.
J Exp Bot
January 2025
Department of Entomology and Plant Pathology, Auburn University, Auburn, AL 36849, USA.
A primary precursor of jasmonates, 12-oxo-phytodienoic acid (OPDA), is an autonomous hormone signal that activates and fine-tunes plant defense responses, as well as growth and development. However, the architecture of its signaling circuits remains largely elusive. Here we describe that OPDA signaling drives photosynthetic reductant powers toward sulfur assimilation in the chloroplasts, incorporating sulfide into cysteine.
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