Substrate-Induced Allosteric Change in the Quaternary Structure of the Spermidine N-Acetyltransferase SpeG
Abstract
The spermidine N-acetyltransferase SpeG is a dodecameric enzyme that catalyzes the transfer of an acetyl group from acetyl coenzyme A to polyamines such as spermidine and spermine. SpeG has an allosteric polyamine-binding site and acetylating polyamines regulate their intracellular concentrations. The structures of SpeG from Vibrio cholerae in complexes with polyamines and cofactor have been characterized earlier. Here, we present the dodecameric structure of SpeG from V. cholerae in a ligand-free form in three different conformational states: open, intermediate and closed. All structures were crystallized in C2 space group symmetry and contain six monomers in the asymmetric unit cell. Two hexamers related by crystallographic 2-fold symmetry form the SpeG dodecamer. The open and intermediate states have a unique open dodecameric ring. This SpeG dodecamer is asymmetric except for the one 2-fold axis and is unlike any known dodecameric structure. Using a fluorescence thermal shift assay, size-exclusion chromatography with multi-angle light scattering, small-angle X-ray scattering analysis, negative-stain electron microscopy and structural analysis, we demonstrate that this unique open dodecameric state exists in solution. As a result, our combined results indicate that polyamines trigger conformational changes and induce the symmetric closed dodecameric state of the protein when they bind to their allostericmore »
- Publication Date:
- Research Org.:
- Argonne National Laboratory (ANL), Argonne, IL (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES)
- OSTI Identifier:
- 1374483
- Alternate Identifier(s):
- OSTI ID: 1248371
- Grant/Contract Number:
- AC02-06CH11357
- Resource Type:
- Published Article
- Journal Name:
- Journal of Molecular Biology
- Additional Journal Information:
- Journal Name: Journal of Molecular Biology Journal Volume: 427 Journal Issue: 22; Journal ID: ISSN 0022-2836
- Publisher:
- Elsevier
- Country of Publication:
- United Kingdom
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; GNAT acetyltransferase; asymmetric structure; allosteric site; dodecameric enzyme; spermidine/spermine
Citation Formats
Filippova, Ekaterina V., Weigand, Steven, Osipiuk, Jerzy, Kiryukhina, Olga, Joachimiak, Andrzej, and Anderson, Wayne F. Substrate-Induced Allosteric Change in the Quaternary Structure of the Spermidine N-Acetyltransferase SpeG. United Kingdom: N. p., 2015.
Web. doi:10.1016/j.jmb.2015.09.013.
Filippova, Ekaterina V., Weigand, Steven, Osipiuk, Jerzy, Kiryukhina, Olga, Joachimiak, Andrzej, & Anderson, Wayne F. Substrate-Induced Allosteric Change in the Quaternary Structure of the Spermidine N-Acetyltransferase SpeG. United Kingdom. https://doi.org/10.1016/j.jmb.2015.09.013
Filippova, Ekaterina V., Weigand, Steven, Osipiuk, Jerzy, Kiryukhina, Olga, Joachimiak, Andrzej, and Anderson, Wayne F. Sun .
"Substrate-Induced Allosteric Change in the Quaternary Structure of the Spermidine N-Acetyltransferase SpeG". United Kingdom. https://doi.org/10.1016/j.jmb.2015.09.013.
@article{osti_1374483,
title = {Substrate-Induced Allosteric Change in the Quaternary Structure of the Spermidine N-Acetyltransferase SpeG},
author = {Filippova, Ekaterina V. and Weigand, Steven and Osipiuk, Jerzy and Kiryukhina, Olga and Joachimiak, Andrzej and Anderson, Wayne F.},
abstractNote = {The spermidine N-acetyltransferase SpeG is a dodecameric enzyme that catalyzes the transfer of an acetyl group from acetyl coenzyme A to polyamines such as spermidine and spermine. SpeG has an allosteric polyamine-binding site and acetylating polyamines regulate their intracellular concentrations. The structures of SpeG from Vibrio cholerae in complexes with polyamines and cofactor have been characterized earlier. Here, we present the dodecameric structure of SpeG from V. cholerae in a ligand-free form in three different conformational states: open, intermediate and closed. All structures were crystallized in C2 space group symmetry and contain six monomers in the asymmetric unit cell. Two hexamers related by crystallographic 2-fold symmetry form the SpeG dodecamer. The open and intermediate states have a unique open dodecameric ring. This SpeG dodecamer is asymmetric except for the one 2-fold axis and is unlike any known dodecameric structure. Using a fluorescence thermal shift assay, size-exclusion chromatography with multi-angle light scattering, small-angle X-ray scattering analysis, negative-stain electron microscopy and structural analysis, we demonstrate that this unique open dodecameric state exists in solution. As a result, our combined results indicate that polyamines trigger conformational changes and induce the symmetric closed dodecameric state of the protein when they bind to their allosteric sites.},
doi = {10.1016/j.jmb.2015.09.013},
journal = {Journal of Molecular Biology},
number = 22,
volume = 427,
place = {United Kingdom},
year = {Sun Nov 01 00:00:00 EDT 2015},
month = {Sun Nov 01 00:00:00 EDT 2015}
}
https://doi.org/10.1016/j.jmb.2015.09.013
Search WorldCat to find libraries that may hold this journalWeb of Science
Works referencing / citing this record:
Analysis of crystalline and solution states of ligand-free spermidine N -acetyltransferase (SpeG) from Escherichia coli
journal, May 2019
- Filippova, Ekaterina V.; Weigand, Steven; Kiryukhina, Olga
- Acta Crystallographica Section D Structural Biology, Vol. 75, Issue 6
The spermidine acetyltransferase SpeG regulates transcription of the small RNA rprA
journal, December 2018
- Hu, Linda I.; Filippova, Ekaterina V.; Dang, Joseph
- PLOS ONE, Vol. 13, Issue 12
Structure and Functional Diversity of GCN5-Related N-Acetyltransferases (GNAT)
journal, June 2016
- Salah Ud-Din, Abu; Tikhomirova, Alexandra; Roujeinikova, Anna
- International Journal of Molecular Sciences, Vol. 17, Issue 7