Structural insights into the hydrolysis and polymorphism of methotrexate polyglutamate by zebrafish γ-Glutamyl hydrolase

Phimonphan Chuankhayan, Tseng Ting Kao, Chien Chih Lin, Hong Hsiang Guan, Atsushi Nakagawa, Tzu-Fun Fu, Chun Jung Chen

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

γ-Glutamyl hydrolases (γGH) catalyze the hydrolysis of γ-linked glutamate residues from the polyglutamyl of folates and antifolates, such as methotrexate (MTX), a widely used anticancer drug. We describe the first crystal structures of the endopeptidase-type γGH (zγGH) from zebrafish and the mutant complexes with MTX(Glu)5 and hydrolyzed MTX(Glu)1, revealing the complete set of key residues involved in hydrolysis as well as the substrate-binding subsites (-1 to +2). The side chain of Phe20 and the 6-methylpterin ring of MTX(Glu)5 invoke π-π interactions to promote distinct concerted conformational alterations involving ∼90 rotations in the complexes with the zγGH-C108A and zγGH-H218N mutant proteins. The structural geometries of the MTX(Glu) 5 and hydrolyzed MTX(Glu)1 in the mutant complexes differ significantly from those of the previously known MTX(Glu)1, providing polymorphic information. Together with the structural comparison and the activity analysis, these results shed light on the catalytic mechanism and substrate recognition of zγGH and other γ-glutamyl hydrolases.

Original languageEnglish
Pages (from-to)7625-7635
Number of pages11
JournalJournal of Medicinal Chemistry
Volume56
Issue number19
DOIs
Publication statusPublished - 2013 Oct 10

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Hydrolases
Zebrafish
Methotrexate
Hydrolysis
Folic Acid Antagonists
Mutant Proteins
methotrexate polyglutamate
Folic Acid
Glutamic Acid
Pharmaceutical Preparations

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Drug Discovery

Cite this

Chuankhayan, Phimonphan ; Kao, Tseng Ting ; Lin, Chien Chih ; Guan, Hong Hsiang ; Nakagawa, Atsushi ; Fu, Tzu-Fun ; Chen, Chun Jung. / Structural insights into the hydrolysis and polymorphism of methotrexate polyglutamate by zebrafish γ-Glutamyl hydrolase. In: Journal of Medicinal Chemistry. 2013 ; Vol. 56, No. 19. pp. 7625-7635.
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abstract = "γ-Glutamyl hydrolases (γGH) catalyze the hydrolysis of γ-linked glutamate residues from the polyglutamyl of folates and antifolates, such as methotrexate (MTX), a widely used anticancer drug. We describe the first crystal structures of the endopeptidase-type γGH (zγGH) from zebrafish and the mutant complexes with MTX(Glu)5 and hydrolyzed MTX(Glu)1, revealing the complete set of key residues involved in hydrolysis as well as the substrate-binding subsites (-1 to +2). The side chain of Phe20 and the 6-methylpterin ring of MTX(Glu)5 invoke π-π interactions to promote distinct concerted conformational alterations involving ∼90 rotations in the complexes with the zγGH-C108A and zγGH-H218N mutant proteins. The structural geometries of the MTX(Glu) 5 and hydrolyzed MTX(Glu)1 in the mutant complexes differ significantly from those of the previously known MTX(Glu)1, providing polymorphic information. Together with the structural comparison and the activity analysis, these results shed light on the catalytic mechanism and substrate recognition of zγGH and other γ-glutamyl hydrolases.",
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Structural insights into the hydrolysis and polymorphism of methotrexate polyglutamate by zebrafish γ-Glutamyl hydrolase. / Chuankhayan, Phimonphan; Kao, Tseng Ting; Lin, Chien Chih; Guan, Hong Hsiang; Nakagawa, Atsushi; Fu, Tzu-Fun; Chen, Chun Jung.

In: Journal of Medicinal Chemistry, Vol. 56, No. 19, 10.10.2013, p. 7625-7635.

Research output: Contribution to journalArticle

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