Abstract
Background: Glutathione transferase (GST) catalyzes glutathione conjugation, a major detoxification pathway for xenobiotics and endogenous substances. Here, we determined the crystal structure of a Delta-class GST from Bombyx mori (bmGSTD) to examine its catalytic residues. Methods: The three-dimensional structure of bmGSTD was resolved by the molecular replacement method and refined to a resolution of 2.0 Å. Results: Structural alignment with a Delta-class GST of Anopheles gambiae indicated that bmGSTD contains 2 distinct domains (an N-terminal domain and a C-terminal domain) connected by a linker. The bound glutathione localized at the N-terminal domain. Putative catalytic residues were changed to alanine by site-directed mutagenesis, and the resulting mutants were characterized in terms of catalytic activity using glutathione and 1-chloro-2,4-dinitrobenzene, a synthetic substrate of GST. Kinetic analysis of bmGSTD mutants indicated that Ser11, Gln51, His52, Ser67, and Arg68 are important for enzyme function. General significance: These results provide structural insights into the catalysis of glutathione conjugation in B. mori by bmGSTD.
| Original language | English |
|---|---|
| Pages (from-to) | 1469-1474 |
| Number of pages | 6 |
| Journal | Biochimica et Biophysica Acta - General Subjects |
| Volume | 1820 |
| Issue number | 10 |
| DOIs | |
| Publication status | Published - Oct 1 2012 |
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All Science Journal Classification (ASJC) codes
- Biophysics
- Biochemistry
- Molecular Biology
Cite this
Structural basis for catalytic activity of a silkworm Delta-class glutathione transferase. / Yamamoto, Kohji; Usuda, Kazuhiro; Kakuta, Yoshimitsu; Kimura, Makoto; Higashiura, Akifumi; Nakagawa, Atsushi; Aso, Yoichi; Suzuki, Mamoru.
In: Biochimica et Biophysica Acta - General Subjects, Vol. 1820, No. 10, 01.10.2012, p. 1469-1474.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Structural basis for catalytic activity of a silkworm Delta-class glutathione transferase
AU - Yamamoto, Kohji
AU - Usuda, Kazuhiro
AU - Kakuta, Yoshimitsu
AU - Kimura, Makoto
AU - Higashiura, Akifumi
AU - Nakagawa, Atsushi
AU - Aso, Yoichi
AU - Suzuki, Mamoru
PY - 2012/10/1
Y1 - 2012/10/1
N2 - Background: Glutathione transferase (GST) catalyzes glutathione conjugation, a major detoxification pathway for xenobiotics and endogenous substances. Here, we determined the crystal structure of a Delta-class GST from Bombyx mori (bmGSTD) to examine its catalytic residues. Methods: The three-dimensional structure of bmGSTD was resolved by the molecular replacement method and refined to a resolution of 2.0 Å. Results: Structural alignment with a Delta-class GST of Anopheles gambiae indicated that bmGSTD contains 2 distinct domains (an N-terminal domain and a C-terminal domain) connected by a linker. The bound glutathione localized at the N-terminal domain. Putative catalytic residues were changed to alanine by site-directed mutagenesis, and the resulting mutants were characterized in terms of catalytic activity using glutathione and 1-chloro-2,4-dinitrobenzene, a synthetic substrate of GST. Kinetic analysis of bmGSTD mutants indicated that Ser11, Gln51, His52, Ser67, and Arg68 are important for enzyme function. General significance: These results provide structural insights into the catalysis of glutathione conjugation in B. mori by bmGSTD.
AB - Background: Glutathione transferase (GST) catalyzes glutathione conjugation, a major detoxification pathway for xenobiotics and endogenous substances. Here, we determined the crystal structure of a Delta-class GST from Bombyx mori (bmGSTD) to examine its catalytic residues. Methods: The three-dimensional structure of bmGSTD was resolved by the molecular replacement method and refined to a resolution of 2.0 Å. Results: Structural alignment with a Delta-class GST of Anopheles gambiae indicated that bmGSTD contains 2 distinct domains (an N-terminal domain and a C-terminal domain) connected by a linker. The bound glutathione localized at the N-terminal domain. Putative catalytic residues were changed to alanine by site-directed mutagenesis, and the resulting mutants were characterized in terms of catalytic activity using glutathione and 1-chloro-2,4-dinitrobenzene, a synthetic substrate of GST. Kinetic analysis of bmGSTD mutants indicated that Ser11, Gln51, His52, Ser67, and Arg68 are important for enzyme function. General significance: These results provide structural insights into the catalysis of glutathione conjugation in B. mori by bmGSTD.
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UR - http://www.scopus.com/inward/citedby.url?scp=84863193817&partnerID=8YFLogxK
U2 - 10.1016/j.bbagen.2012.04.022
DO - 10.1016/j.bbagen.2012.04.022
M3 - Article
C2 - 22579926
AN - SCOPUS:84863193817
VL - 1820
SP - 1469
EP - 1474
JO - Biochimica et Biophysica Acta - General Subjects
JF - Biochimica et Biophysica Acta - General Subjects
SN - 0304-4165
IS - 10
ER -