Biochemical and structural characterization of 5′-methylthioadenosine nucleosidases from Arabidopsis thaliana

Eun Young Park; Woo Suk Choi; Seung Ick Oh; Kyung Nam Kim; Jeong Sheop Shin; Hyun Kyu Song

doi:10.1016/j.bbrc.2009.02.106

Biochemical and structural characterization of 5′-methylthioadenosine nucleosidases from Arabidopsis thaliana

Eun Young Park, Woo Suk Choi, Seung Ick Oh, Kyung Nam Kim, Jeong Sheop Shin, Hyun Kyu Song

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

6 Citations (Scopus)

Abstract

5′-Methylthioadenosine (MTA) and S-adenosylhomocysteine (SAH) are important metabolites in all living organisms. Two similar nucleosidases for hydrolyzing MTA in Arabidopsis thaliana (AtMTAN1 and AtMTAN2) exist, but only AtMTAN2 shows markedly broad substrate specificity for hydrolysis of SAH. To examine the biochemical characteristics of AtMTAN2, it was over-expressed in Escherichia coli and purified to homogeneity. Spectroscopic assays confirm AtMTAN2 catalyzes MTA as well as SAH hydrolysis, compared to AtMTAN1 which only hydrolyzes MTA. In addition, crystal structure of the AtMTAN2 enzyme in complex with, adenine was determined at 2.9 Å resolution. Finally, a structural comparison of AtMTAN2 performed with previously determined structures of AtMTAN1 and an E. coli homolog provides clues for the substrate specificity of MTA nucleosidases in A. thaliana.

Original language	English
Pages (from-to)	619-624
Number of pages	6
Journal	Biochemical and biophysical research communications
Volume	381
Issue number	4
DOIs	https://doi.org/10.1016/j.bbrc.2009.02.106
Publication status	Published - 2009 Apr 17

Bibliographical note

Funding Information:
We thank the staff at 4A beamline, Pohang Light Source, Korea and NW12 beamline, Photon Factory, Japan for helping with the data collection. This work was supported by grants from the Plant Signaling Network Research Center and Molecular and Cellular Biodiscovery Research Program (M10648230002-08N4823-00210), Korea Science and Engineering Foundation.

Keywords

Adenine
Crystal structure
Flexible loop
MTA
Nucleosidase
Plant
SAH
Spectroscopic assay

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/j.bbrc.2009.02.106

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title = "Biochemical and structural characterization of 5′-methylthioadenosine nucleosidases from Arabidopsis thaliana",

abstract = "5′-Methylthioadenosine (MTA) and S-adenosylhomocysteine (SAH) are important metabolites in all living organisms. Two similar nucleosidases for hydrolyzing MTA in Arabidopsis thaliana (AtMTAN1 and AtMTAN2) exist, but only AtMTAN2 shows markedly broad substrate specificity for hydrolysis of SAH. To examine the biochemical characteristics of AtMTAN2, it was over-expressed in Escherichia coli and purified to homogeneity. Spectroscopic assays confirm AtMTAN2 catalyzes MTA as well as SAH hydrolysis, compared to AtMTAN1 which only hydrolyzes MTA. In addition, crystal structure of the AtMTAN2 enzyme in complex with, adenine was determined at 2.9 {\AA} resolution. Finally, a structural comparison of AtMTAN2 performed with previously determined structures of AtMTAN1 and an E. coli homolog provides clues for the substrate specificity of MTA nucleosidases in A. thaliana.",

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note = "Funding Information: We thank the staff at 4A beamline, Pohang Light Source, Korea and NW12 beamline, Photon Factory, Japan for helping with the data collection. This work was supported by grants from the Plant Signaling Network Research Center and Molecular and Cellular Biodiscovery Research Program (M10648230002-08N4823-00210), Korea Science and Engineering Foundation. ",

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AU - Park, Eun Young

AU - Choi, Woo Suk

AU - Oh, Seung Ick

AU - Kim, Kyung Nam

AU - Shin, Jeong Sheop

AU - Song, Hyun Kyu

N1 - Funding Information: We thank the staff at 4A beamline, Pohang Light Source, Korea and NW12 beamline, Photon Factory, Japan for helping with the data collection. This work was supported by grants from the Plant Signaling Network Research Center and Molecular and Cellular Biodiscovery Research Program (M10648230002-08N4823-00210), Korea Science and Engineering Foundation.

PY - 2009/4/17

Y1 - 2009/4/17

N2 - 5′-Methylthioadenosine (MTA) and S-adenosylhomocysteine (SAH) are important metabolites in all living organisms. Two similar nucleosidases for hydrolyzing MTA in Arabidopsis thaliana (AtMTAN1 and AtMTAN2) exist, but only AtMTAN2 shows markedly broad substrate specificity for hydrolysis of SAH. To examine the biochemical characteristics of AtMTAN2, it was over-expressed in Escherichia coli and purified to homogeneity. Spectroscopic assays confirm AtMTAN2 catalyzes MTA as well as SAH hydrolysis, compared to AtMTAN1 which only hydrolyzes MTA. In addition, crystal structure of the AtMTAN2 enzyme in complex with, adenine was determined at 2.9 Å resolution. Finally, a structural comparison of AtMTAN2 performed with previously determined structures of AtMTAN1 and an E. coli homolog provides clues for the substrate specificity of MTA nucleosidases in A. thaliana.

AB - 5′-Methylthioadenosine (MTA) and S-adenosylhomocysteine (SAH) are important metabolites in all living organisms. Two similar nucleosidases for hydrolyzing MTA in Arabidopsis thaliana (AtMTAN1 and AtMTAN2) exist, but only AtMTAN2 shows markedly broad substrate specificity for hydrolysis of SAH. To examine the biochemical characteristics of AtMTAN2, it was over-expressed in Escherichia coli and purified to homogeneity. Spectroscopic assays confirm AtMTAN2 catalyzes MTA as well as SAH hydrolysis, compared to AtMTAN1 which only hydrolyzes MTA. In addition, crystal structure of the AtMTAN2 enzyme in complex with, adenine was determined at 2.9 Å resolution. Finally, a structural comparison of AtMTAN2 performed with previously determined structures of AtMTAN1 and an E. coli homolog provides clues for the substrate specificity of MTA nucleosidases in A. thaliana.

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KW - Flexible loop

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KW - Nucleosidase

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Biochemical and structural characterization of 5′-methylthioadenosine nucleosidases from Arabidopsis thaliana

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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