Binding and inhibition of human spermidine synthase by decarboxylated S-adenosylhomocysteine

Jolita Šečkute, Diane E. McCloskey, H. Jeanette Thomas, John A. Secrist, Anthony Pegg, Steven E. Ealick

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Aminopropyltransferases are essential enzymes that form polyamines in eukaryotic and most prokaryotic cells. Spermidine synthase (SpdS) is one of the most well-studied enzymes in this biosynthetic pathway. The enzyme uses decarboxylated S-adenosylmethionine and a short-chain polyamine (putrescine) to make a medium-chain polyamine (spermidine) and 5′-deoxy-5′- methylthioadenosine as a byproduct. Here, we report a new spermidine synthase inhibitor, decarboxylated S-adenosylhomocysteine (dcSAH). The inhibitor was synthesized, and dosedependent inhibition of human, Thermatoga maritima, and Plasmodium falciparum spermidine synthases, as well as functionally homologous human spermine synthase, was determined. The human SpdS/dcSAH complex structure was determined by X-ray crystallography at 2.0 Å resolution and showed consistent active site positioning and coordination with previously known structures. Isothermal calorimetry binding assays confirmed inhibitor binding to human SpdS with K d of 1.1 ± 0.3 μM in the absence of putrescine and 3.2 ± 0.1 lM in the presence of putrescine. These results indicate a potential for further inhibitor development based on the dcSAH scaffold. Published by Wiley-Blackwell.

Original languageEnglish (US)
Pages (from-to)1836-1844
Number of pages9
JournalProtein Science
Volume20
Issue number11
DOIs
StatePublished - Nov 1 2011

Fingerprint

Spermidine Synthase
Putrescine
Polyamines
Spermine Synthase
Enzymes
Prokaryotic Cells
Calorimetry
Spermidine
X ray crystallography
X Ray Crystallography
Biosynthetic Pathways
Plasmodium falciparum
Scaffolds
Byproducts
S-adenosyl-3-thiopropylamine
Assays
Catalytic Domain

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Šečkute, J., McCloskey, D. E., Thomas, H. J., Secrist, J. A., Pegg, A., & Ealick, S. E. (2011). Binding and inhibition of human spermidine synthase by decarboxylated S-adenosylhomocysteine. Protein Science, 20(11), 1836-1844. https://doi.org/10.1002/pro.717
Šečkute, Jolita ; McCloskey, Diane E. ; Thomas, H. Jeanette ; Secrist, John A. ; Pegg, Anthony ; Ealick, Steven E. / Binding and inhibition of human spermidine synthase by decarboxylated S-adenosylhomocysteine. In: Protein Science. 2011 ; Vol. 20, No. 11. pp. 1836-1844.
@article{03ff9fa661ff4007853c3dc4af3c5e94,
title = "Binding and inhibition of human spermidine synthase by decarboxylated S-adenosylhomocysteine",
abstract = "Aminopropyltransferases are essential enzymes that form polyamines in eukaryotic and most prokaryotic cells. Spermidine synthase (SpdS) is one of the most well-studied enzymes in this biosynthetic pathway. The enzyme uses decarboxylated S-adenosylmethionine and a short-chain polyamine (putrescine) to make a medium-chain polyamine (spermidine) and 5′-deoxy-5′- methylthioadenosine as a byproduct. Here, we report a new spermidine synthase inhibitor, decarboxylated S-adenosylhomocysteine (dcSAH). The inhibitor was synthesized, and dosedependent inhibition of human, Thermatoga maritima, and Plasmodium falciparum spermidine synthases, as well as functionally homologous human spermine synthase, was determined. The human SpdS/dcSAH complex structure was determined by X-ray crystallography at 2.0 {\AA} resolution and showed consistent active site positioning and coordination with previously known structures. Isothermal calorimetry binding assays confirmed inhibitor binding to human SpdS with K d of 1.1 ± 0.3 μM in the absence of putrescine and 3.2 ± 0.1 lM in the presence of putrescine. These results indicate a potential for further inhibitor development based on the dcSAH scaffold. Published by Wiley-Blackwell.",
author = "Jolita Šečkute and McCloskey, {Diane E.} and Thomas, {H. Jeanette} and Secrist, {John A.} and Anthony Pegg and Ealick, {Steven E.}",
year = "2011",
month = "11",
day = "1",
doi = "10.1002/pro.717",
language = "English (US)",
volume = "20",
pages = "1836--1844",
journal = "Protein Science",
issn = "0961-8368",
publisher = "Cold Spring Harbor Laboratory Press",
number = "11",

}

Šečkute, J, McCloskey, DE, Thomas, HJ, Secrist, JA, Pegg, A & Ealick, SE 2011, 'Binding and inhibition of human spermidine synthase by decarboxylated S-adenosylhomocysteine', Protein Science, vol. 20, no. 11, pp. 1836-1844. https://doi.org/10.1002/pro.717

Binding and inhibition of human spermidine synthase by decarboxylated S-adenosylhomocysteine. / Šečkute, Jolita; McCloskey, Diane E.; Thomas, H. Jeanette; Secrist, John A.; Pegg, Anthony; Ealick, Steven E.

In: Protein Science, Vol. 20, No. 11, 01.11.2011, p. 1836-1844.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Binding and inhibition of human spermidine synthase by decarboxylated S-adenosylhomocysteine

AU - Šečkute, Jolita

AU - McCloskey, Diane E.

AU - Thomas, H. Jeanette

AU - Secrist, John A.

AU - Pegg, Anthony

AU - Ealick, Steven E.

PY - 2011/11/1

Y1 - 2011/11/1

N2 - Aminopropyltransferases are essential enzymes that form polyamines in eukaryotic and most prokaryotic cells. Spermidine synthase (SpdS) is one of the most well-studied enzymes in this biosynthetic pathway. The enzyme uses decarboxylated S-adenosylmethionine and a short-chain polyamine (putrescine) to make a medium-chain polyamine (spermidine) and 5′-deoxy-5′- methylthioadenosine as a byproduct. Here, we report a new spermidine synthase inhibitor, decarboxylated S-adenosylhomocysteine (dcSAH). The inhibitor was synthesized, and dosedependent inhibition of human, Thermatoga maritima, and Plasmodium falciparum spermidine synthases, as well as functionally homologous human spermine synthase, was determined. The human SpdS/dcSAH complex structure was determined by X-ray crystallography at 2.0 Å resolution and showed consistent active site positioning and coordination with previously known structures. Isothermal calorimetry binding assays confirmed inhibitor binding to human SpdS with K d of 1.1 ± 0.3 μM in the absence of putrescine and 3.2 ± 0.1 lM in the presence of putrescine. These results indicate a potential for further inhibitor development based on the dcSAH scaffold. Published by Wiley-Blackwell.

AB - Aminopropyltransferases are essential enzymes that form polyamines in eukaryotic and most prokaryotic cells. Spermidine synthase (SpdS) is one of the most well-studied enzymes in this biosynthetic pathway. The enzyme uses decarboxylated S-adenosylmethionine and a short-chain polyamine (putrescine) to make a medium-chain polyamine (spermidine) and 5′-deoxy-5′- methylthioadenosine as a byproduct. Here, we report a new spermidine synthase inhibitor, decarboxylated S-adenosylhomocysteine (dcSAH). The inhibitor was synthesized, and dosedependent inhibition of human, Thermatoga maritima, and Plasmodium falciparum spermidine synthases, as well as functionally homologous human spermine synthase, was determined. The human SpdS/dcSAH complex structure was determined by X-ray crystallography at 2.0 Å resolution and showed consistent active site positioning and coordination with previously known structures. Isothermal calorimetry binding assays confirmed inhibitor binding to human SpdS with K d of 1.1 ± 0.3 μM in the absence of putrescine and 3.2 ± 0.1 lM in the presence of putrescine. These results indicate a potential for further inhibitor development based on the dcSAH scaffold. Published by Wiley-Blackwell.

UR - http://www.scopus.com/inward/record.url?scp=80054753541&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=80054753541&partnerID=8YFLogxK

U2 - 10.1002/pro.717

DO - 10.1002/pro.717

M3 - Article

C2 - 21898642

AN - SCOPUS:80054753541

VL - 20

SP - 1836

EP - 1844

JO - Protein Science

JF - Protein Science

SN - 0961-8368

IS - 11

ER -