Analysis of crystalline and solution states of ligand-free spermidine N-acetyltransferase (SpeG) from escherichia coli

Ekaterina V. Filippova; Steven Weigand; Olga Kiryukhina; Alan J. Wolfe; Wayne F. Anderson

doi:10.1107/S2059798319006545

Analysis of crystalline and solution states of ligand-free spermidine N-acetyltransferase (SpeG) from escherichia coli

Ekaterina V. Filippova^*, Steven Weigand, Olga Kiryukhina, Alan J. Wolfe, Wayne F. Anderson

^*Corresponding author for this work

Biochemistry and Molecular Genetics

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

Spermidine N-acetyltransferase (SpeG) transfers an acetyl group from acetylcoenzyme A to an N-terminal amino group of intracellular spermidine. This acetylation inactivates spermidine, reducing the polyamine toxicity that tends to occur under certain chemical and physical stresses. The structure of the SpeG protein from Vibrio cholerae has been characterized: while the monomer possesses a structural fold similar to those of other Gcn5-related N-acetyltransferase superfamily members, its dodecameric structure remains exceptional. In this paper, structural analyses of SpeG isolated from Escherichia coli are described. Like V. cholerae SpeG, E. coli SpeG forms dodecamers, as revealed by two crystal structures of the ligand-free E. coli SpeG dodecamer determined at 1.75 and 2.9 Å resolution. Although both V. cholerae SpeG and E. coli SpeG can adopt an asymmetric open dodecameric state, solution analysis showed that the oligomeric composition of ligand-free E. coli SpeG differs from that of ligand-free V. cholerae SpeG. Based on these data, it is proposed that the equilibrium balance of SpeG oligomers in the absence of ligands differs from one species to another and thus might be important for SpeG function.

Original language	English (US)
Pages (from-to)	545-553
Number of pages	9
Journal	Acta Crystallographica Section D: Structural Biology
Volume	75
DOIs	https://doi.org/10.1107/S2059798319006545
State	Published - Jun 1 2019

Funding

This research was supported in part by the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services under Contract Nos. HHSN272200700058C, HHSN272201200026C and HHSN272201700060C (WFA). The X-ray and SAXS data collection was performed at the LS-CAT Sector 21 and DND-CAT Sector 5 at the Advanced Photon Source supported by the Argonne National Laboratory operated by the University of Chicago Argonne LLC for the US Department of Energy, Office of Biological and Environmental Research under contract DE-AC0206CH11357. We thank the Keck Biophysics Facility at Northwestern University, Evanston, Illinois, USA for assistance with the SEC-MALS experiment. The negative-staining EM experiment was performed at the Structural Biology Facility at Northwestern University. The LS-CAT is supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor (Grant 085P1000817). The EM research was supported in part by the Searle Leadership Fund for the Life Sciences at Northwestern University, established by the Searle Funds at The Chicago Community Trust. This research was supported in part by the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services under Contract Nos. HHSN272200700058C, HHSN272201200026C and HHSN272201700060C (WFA). The X-ray and SAXS data collection was performed at the LS-CAT Sector 21 and DND-CAT Sector 5 at the Advanced Photon Source supported by the Argonne National Laboratory operated by the University of Chicago Argonne LLC for the US Department of Energy, Office of Biological and Environmental Research under contract DE-AC02-06CH11357. We thank the Keck Biophysics Facility at Northwestern University, Evanston, Illinois, USA for assistance with the SEC-MALS experiment. The negative-staining EM experiment was performed at the Structural Biology Facility at Northwestern University. The LS-CAT is supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor (Grant 085P1000817). The EM research was supported in part by the Searle Leadership Fund for the Life Sciences at Northwestern University, established by the Searle Funds at The Chicago Community Trust.

Keywords

Escherichia coli
GNAT family
Polyamine acetylation
SpeG
Spermidine N-acetyltransferase

ASJC Scopus subject areas

Structural Biology

Access to Document

10.1107/S2059798319006545

Cite this

@article{42b1300328954823aee701b610d52b67,

title = "Analysis of crystalline and solution states of ligand-free spermidine N-acetyltransferase (SpeG) from escherichia coli",

abstract = "Spermidine N-acetyltransferase (SpeG) transfers an acetyl group from acetylcoenzyme A to an N-terminal amino group of intracellular spermidine. This acetylation inactivates spermidine, reducing the polyamine toxicity that tends to occur under certain chemical and physical stresses. The structure of the SpeG protein from Vibrio cholerae has been characterized: while the monomer possesses a structural fold similar to those of other Gcn5-related N-acetyltransferase superfamily members, its dodecameric structure remains exceptional. In this paper, structural analyses of SpeG isolated from Escherichia coli are described. Like V. cholerae SpeG, E. coli SpeG forms dodecamers, as revealed by two crystal structures of the ligand-free E. coli SpeG dodecamer determined at 1.75 and 2.9 {\AA} resolution. Although both V. cholerae SpeG and E. coli SpeG can adopt an asymmetric open dodecameric state, solution analysis showed that the oligomeric composition of ligand-free E. coli SpeG differs from that of ligand-free V. cholerae SpeG. Based on these data, it is proposed that the equilibrium balance of SpeG oligomers in the absence of ligands differs from one species to another and thus might be important for SpeG function.",

keywords = "Escherichia coli, GNAT family, Polyamine acetylation, SpeG, Spermidine N-acetyltransferase",

author = "Filippova, {Ekaterina V.} and Steven Weigand and Olga Kiryukhina and Wolfe, {Alan J.} and Anderson, {Wayne F.}",

note = "Funding Information: This research was supported in part by the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services under Contract Nos. HHSN272200700058C, HHSN272201200026C and HHSN272201700060C (WFA). Funding Information: The X-ray and SAXS data collection was performed at the LS-CAT Sector 21 and DND-CAT Sector 5 at the Advanced Photon Source supported by the Argonne National Laboratory operated by the University of Chicago Argonne LLC for the US Department of Energy, Office of Biological and Environmental Research under contract DE-AC0206CH11357. We thank the Keck Biophysics Facility at Northwestern University, Evanston, Illinois, USA for assistance with the SEC-MALS experiment. The negative-staining EM experiment was performed at the Structural Biology Facility at Northwestern University. The LS-CAT is supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor (Grant 085P1000817). The EM research was supported in part by the Searle Leadership Fund for the Life Sciences at Northwestern University, established by the Searle Funds at The Chicago Community Trust. This research was supported in part by the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services under Contract Nos. HHSN272200700058C, HHSN272201200026C and HHSN272201700060C (WFA). Funding Information: The X-ray and SAXS data collection was performed at the LS-CAT Sector 21 and DND-CAT Sector 5 at the Advanced Photon Source supported by the Argonne National Laboratory operated by the University of Chicago Argonne LLC for the US Department of Energy, Office of Biological and Environmental Research under contract DE-AC02-06CH11357. We thank the Keck Biophysics Facility at Northwestern University, Evanston, Illinois, USA for assistance with the SEC-MALS experiment. The negative-staining EM experiment was performed at the Structural Biology Facility at Northwestern University. The LS-CAT is supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor (Grant 085P1000817). The EM research was supported in part by the Searle Leadership Fund for the Life Sciences at Northwestern University, established by the Searle Funds at The Chicago Community Trust. Publisher Copyright: {\textcopyright} 2019 International Union of Crystallography.",

year = "2019",

month = jun,

day = "1",

doi = "10.1107/S2059798319006545",

language = "English (US)",

volume = "75",

pages = "545--553",

journal = "Acta Crystallographica Section D: Structural Biology",

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publisher = "International Union of Crystallography",

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TY - JOUR

T1 - Analysis of crystalline and solution states of ligand-free spermidine N-acetyltransferase (SpeG) from escherichia coli

AU - Filippova, Ekaterina V.

AU - Weigand, Steven

AU - Kiryukhina, Olga

AU - Wolfe, Alan J.

AU - Anderson, Wayne F.

N1 - Funding Information: This research was supported in part by the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services under Contract Nos. HHSN272200700058C, HHSN272201200026C and HHSN272201700060C (WFA). Funding Information: The X-ray and SAXS data collection was performed at the LS-CAT Sector 21 and DND-CAT Sector 5 at the Advanced Photon Source supported by the Argonne National Laboratory operated by the University of Chicago Argonne LLC for the US Department of Energy, Office of Biological and Environmental Research under contract DE-AC0206CH11357. We thank the Keck Biophysics Facility at Northwestern University, Evanston, Illinois, USA for assistance with the SEC-MALS experiment. The negative-staining EM experiment was performed at the Structural Biology Facility at Northwestern University. The LS-CAT is supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor (Grant 085P1000817). The EM research was supported in part by the Searle Leadership Fund for the Life Sciences at Northwestern University, established by the Searle Funds at The Chicago Community Trust. This research was supported in part by the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services under Contract Nos. HHSN272200700058C, HHSN272201200026C and HHSN272201700060C (WFA). Funding Information: The X-ray and SAXS data collection was performed at the LS-CAT Sector 21 and DND-CAT Sector 5 at the Advanced Photon Source supported by the Argonne National Laboratory operated by the University of Chicago Argonne LLC for the US Department of Energy, Office of Biological and Environmental Research under contract DE-AC02-06CH11357. We thank the Keck Biophysics Facility at Northwestern University, Evanston, Illinois, USA for assistance with the SEC-MALS experiment. The negative-staining EM experiment was performed at the Structural Biology Facility at Northwestern University. The LS-CAT is supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor (Grant 085P1000817). The EM research was supported in part by the Searle Leadership Fund for the Life Sciences at Northwestern University, established by the Searle Funds at The Chicago Community Trust. Publisher Copyright: © 2019 International Union of Crystallography.

PY - 2019/6/1

Y1 - 2019/6/1

N2 - Spermidine N-acetyltransferase (SpeG) transfers an acetyl group from acetylcoenzyme A to an N-terminal amino group of intracellular spermidine. This acetylation inactivates spermidine, reducing the polyamine toxicity that tends to occur under certain chemical and physical stresses. The structure of the SpeG protein from Vibrio cholerae has been characterized: while the monomer possesses a structural fold similar to those of other Gcn5-related N-acetyltransferase superfamily members, its dodecameric structure remains exceptional. In this paper, structural analyses of SpeG isolated from Escherichia coli are described. Like V. cholerae SpeG, E. coli SpeG forms dodecamers, as revealed by two crystal structures of the ligand-free E. coli SpeG dodecamer determined at 1.75 and 2.9 Å resolution. Although both V. cholerae SpeG and E. coli SpeG can adopt an asymmetric open dodecameric state, solution analysis showed that the oligomeric composition of ligand-free E. coli SpeG differs from that of ligand-free V. cholerae SpeG. Based on these data, it is proposed that the equilibrium balance of SpeG oligomers in the absence of ligands differs from one species to another and thus might be important for SpeG function.

AB - Spermidine N-acetyltransferase (SpeG) transfers an acetyl group from acetylcoenzyme A to an N-terminal amino group of intracellular spermidine. This acetylation inactivates spermidine, reducing the polyamine toxicity that tends to occur under certain chemical and physical stresses. The structure of the SpeG protein from Vibrio cholerae has been characterized: while the monomer possesses a structural fold similar to those of other Gcn5-related N-acetyltransferase superfamily members, its dodecameric structure remains exceptional. In this paper, structural analyses of SpeG isolated from Escherichia coli are described. Like V. cholerae SpeG, E. coli SpeG forms dodecamers, as revealed by two crystal structures of the ligand-free E. coli SpeG dodecamer determined at 1.75 and 2.9 Å resolution. Although both V. cholerae SpeG and E. coli SpeG can adopt an asymmetric open dodecameric state, solution analysis showed that the oligomeric composition of ligand-free E. coli SpeG differs from that of ligand-free V. cholerae SpeG. Based on these data, it is proposed that the equilibrium balance of SpeG oligomers in the absence of ligands differs from one species to another and thus might be important for SpeG function.

KW - Escherichia coli

KW - GNAT family

KW - Polyamine acetylation

KW - SpeG

KW - Spermidine N-acetyltransferase

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Analysis of crystalline and solution states of ligand-free spermidine N-acetyltransferase (SpeG) from escherichia coli

Abstract

Funding

Keywords

ASJC Scopus subject areas

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Crystal structure of spermidine/spermine N-acetyltransferase SpeG from Escherichia coli in complex with tris(hydroxymethyl)aminomethane.

Crystal structure of spermidine N-acetyltransferase from Escherichia coli

Cite this

Analysis of crystalline and solution states of ligand-free spermidine N-acetyltransferase (SpeG) from escherichia coli

Abstract

Funding

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Datasets

Crystal structure of spermidine/spermine N-acetyltransferase SpeG from Escherichia coli in complex with tris(hydroxymethyl)aminomethane.

Crystal structure of spermidine N-acetyltransferase from Escherichia coli

Cite this