TY - JOUR
T1 - Crystal structure of a Bombyx mori sigma-class glutathione transferase exhibiting prostaglandin e synthase activity
AU - Yamamoto, Kohji
AU - Higashiura, Akifumi
AU - Suzuki, Mamoru
AU - Aritake, Kosuke
AU - Urade, Yoshihiro
AU - Uodome, Nobuko
AU - Nakagawa, Atsushi
N1 - Funding Information:
This work was supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan . The work was also supported in part by the Research Grant for Young Investigators of Faculty of Agriculture, Kyushu University . This work was performed under the Cooperative Research Program of Institute for Protein Research, Osaka University . The synchrotron radiation experiments were carried out at the BL44XU of SPring-8 with the approval of the Japan Synchrotron Radiation Research Institute (JASRI) (Proposal No. 2011A6651 , and 2011B6651 ).
Copyright:
Copyright 2013 Elsevier B.V., All rights reserved.
PY - 2013/6
Y1 - 2013/6
N2 - Background Glutathione transferases (GSTs) are members of a major family of detoxification enzymes. Here, we report the crystal structure of a sigma-class GST of Bombyx mori, bmGSTS1, to gain insight into the mechanism catalysis. Methods The structure of bmGSTS1 and its complex with glutathione were determined at resolutions of 1.9 Å and 1.7 Å by synchrotron radiation and the molecular replacement method. Results The three-dimensional structure of bmGSTS1 shows that it exists as a dimer and is similar in structure to other GSTs with respect to its secondary and tertiary structures. Although striking similarities to the structure of prostaglandin D synthase were also detected, we were surprised to find that bmGSTS1 can convert prostaglandin H2 into its E2 form. Comparison of bmGSTS1 with its glutathione complex showed that bound glutathione was localized to the glutathione-binding site (G-site). Site-directed mutagenesis of bmGSTS1 mutants indicated that amino acid residues Tyr8, Leu14, Trp39, Lys43, Gln50, Met51, Gln63, and Ser64 in the G-site contribute to catalytic activity. Conclusion We determined the tertiary structure of bmGSTS1 exhibiting prostaglandin E synthase activity. General significance These results are, to our knowledge, the first report of a prostaglandin synthase activity in insects.
AB - Background Glutathione transferases (GSTs) are members of a major family of detoxification enzymes. Here, we report the crystal structure of a sigma-class GST of Bombyx mori, bmGSTS1, to gain insight into the mechanism catalysis. Methods The structure of bmGSTS1 and its complex with glutathione were determined at resolutions of 1.9 Å and 1.7 Å by synchrotron radiation and the molecular replacement method. Results The three-dimensional structure of bmGSTS1 shows that it exists as a dimer and is similar in structure to other GSTs with respect to its secondary and tertiary structures. Although striking similarities to the structure of prostaglandin D synthase were also detected, we were surprised to find that bmGSTS1 can convert prostaglandin H2 into its E2 form. Comparison of bmGSTS1 with its glutathione complex showed that bound glutathione was localized to the glutathione-binding site (G-site). Site-directed mutagenesis of bmGSTS1 mutants indicated that amino acid residues Tyr8, Leu14, Trp39, Lys43, Gln50, Met51, Gln63, and Ser64 in the G-site contribute to catalytic activity. Conclusion We determined the tertiary structure of bmGSTS1 exhibiting prostaglandin E synthase activity. General significance These results are, to our knowledge, the first report of a prostaglandin synthase activity in insects.
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U2 - 10.1016/j.bbagen.2013.02.021
DO - 10.1016/j.bbagen.2013.02.021
M3 - Article
C2 - 23458683
AN - SCOPUS:84876008796
VL - 1830
SP - 3711
EP - 3718
JO - Biochimica et Biophysica Acta - General Subjects
JF - Biochimica et Biophysica Acta - General Subjects
SN - 0304-4165
IS - 6
ER -