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Title: Intrinsic disorder in the regulatory N-terminal domain of diacylglycerol acyltransferase 1 from Brassica napus

Abstract

Proteins with multifunctional regulatory domains often demonstrate structural plasticity or protein disorder, allowing the binding of multiple regulatory factors and post-translational modifications. While the importance of protein disorder is clear, it also poses a challenge for in vitro characterization. Here, we report protein intrinsic disorder in a plant molecular system, which despite its prevalence is less studied. We present a detailed biophysical characterization of the entire cytoplasmic N-terminal domain of Brassica napus diacylglycerol acyltransferase, (DGAT1), which includes an inhibitory module and allosteric binding sites. Our results demonstrate that the monomeric N-terminal domain can be stabilized for biophysical characterization and is largely intrinsically disordered in solution. This domain interacts with allosteric modulators of DGAT1, CoA and oleoyl-CoA, at micromolar concentrations. Furthermore, while solution scattering studies indicate conformational heterogeneity in the N-terminal domain of DGAT1, there is a small gain of secondary structure induced by ligand binding.

Authors:
 [1];  [2];  [1];  [1];  [1];  [1];  [1]
  1. Univ. of Alberta, Edmonton, AB (Canada)
  2. Stanford Univ., Menlo Park, CA (United States)
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1490671
Grant/Contract Number:  
AC02-76SF00515
Resource Type:
Accepted Manuscript
Journal Name:
Scientific Reports
Additional Journal Information:
Journal Volume: 8; Journal Issue: 1; Journal ID: ISSN 2045-2322
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Panigrahi, Rashmi, Matsui, Tsutomu, Song, Andrew H., Caldo, Kristian Mark P., Young, Howard S., Weselake, Randall J., and Lemieux, M. Joanne. Intrinsic disorder in the regulatory N-terminal domain of diacylglycerol acyltransferase 1 from Brassica napus. United States: N. p., 2018. Web. doi:10.1038/s41598-018-34339-1.
Panigrahi, Rashmi, Matsui, Tsutomu, Song, Andrew H., Caldo, Kristian Mark P., Young, Howard S., Weselake, Randall J., & Lemieux, M. Joanne. Intrinsic disorder in the regulatory N-terminal domain of diacylglycerol acyltransferase 1 from Brassica napus. United States. doi:10.1038/s41598-018-34339-1.
Panigrahi, Rashmi, Matsui, Tsutomu, Song, Andrew H., Caldo, Kristian Mark P., Young, Howard S., Weselake, Randall J., and Lemieux, M. Joanne. Mon . "Intrinsic disorder in the regulatory N-terminal domain of diacylglycerol acyltransferase 1 from Brassica napus". United States. doi:10.1038/s41598-018-34339-1. https://www.osti.gov/servlets/purl/1490671.
@article{osti_1490671,
title = {Intrinsic disorder in the regulatory N-terminal domain of diacylglycerol acyltransferase 1 from Brassica napus},
author = {Panigrahi, Rashmi and Matsui, Tsutomu and Song, Andrew H. and Caldo, Kristian Mark P. and Young, Howard S. and Weselake, Randall J. and Lemieux, M. Joanne},
abstractNote = {Proteins with multifunctional regulatory domains often demonstrate structural plasticity or protein disorder, allowing the binding of multiple regulatory factors and post-translational modifications. While the importance of protein disorder is clear, it also poses a challenge for in vitro characterization. Here, we report protein intrinsic disorder in a plant molecular system, which despite its prevalence is less studied. We present a detailed biophysical characterization of the entire cytoplasmic N-terminal domain of Brassica napus diacylglycerol acyltransferase, (DGAT1), which includes an inhibitory module and allosteric binding sites. Our results demonstrate that the monomeric N-terminal domain can be stabilized for biophysical characterization and is largely intrinsically disordered in solution. This domain interacts with allosteric modulators of DGAT1, CoA and oleoyl-CoA, at micromolar concentrations. Furthermore, while solution scattering studies indicate conformational heterogeneity in the N-terminal domain of DGAT1, there is a small gain of secondary structure induced by ligand binding.},
doi = {10.1038/s41598-018-34339-1},
journal = {Scientific Reports},
number = 1,
volume = 8,
place = {United States},
year = {2018},
month = {11}
}

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