Details
| Original language | English |
|---|---|
| Article number | 4160 |
| Journal | Scientific reports |
| Volume | 7 |
| Issue number | 1 |
| Publication status | Published - 1 Dec 2017 |
Abstract
Sulphotransferases are a diverse group of enzymes catalysing the transfer of a sulfuryl group from 3′-phosphoadenosine 5′-phosphosulphate (PAPS) to a broad range of secondary metabolites. They exist in all kingdoms of life. In Arabidopsis thaliana (L.) Heynh. twenty-two sulphotransferase (SOT) isoforms were identified. Three of those are involved in glucosinolate (Gl) biosynthesis, glycosylated sulphur-containing aldoximes containing chemically different side chains, whose break-down products are involved in stress response against herbivores, pathogens, and abiotic stress. To explain the differences in substrate specificity of desulpho (ds)-Gl SOTs and to understand the reaction mechanism of plant SOTs, we determined the first high-resolution crystal structure of the plant ds-Gl SOT AtSOT18 in complex with 3′-phosphoadenosine 5′-phosphate (PAP) alone and together with the Gl sinigrin. These new structural insights into the determination of substrate specificity were complemented by mutagenesis studies. The structure of AtSOT18 invigorates the similarity between plant and mammalian sulphotransferases, which illustrates the evolutionary conservation of this multifunctional enzyme family. We identified the essential residues for substrate binding and catalysis and demonstrated that the catalytic mechanism is conserved between human and plant enzymes. Our study indicates that the loop-gating mechanism is likely to be a source of the substrate specificity in plants.
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In: Scientific reports, Vol. 7, No. 1, 4160, 01.12.2017.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Structural and biochemical studies of sulphotransferase 18 from Arabidopsis thaliana explain its substrate specificity and reaction mechanism
AU - Hirschmann, Felix
AU - Baruch, Petra
AU - Chizhov, Igor
AU - Mueller, Jonathan Wolf
AU - Manstein, Dietmar J.
AU - Papenbrock, Jutta
AU - Fedorov, Roman
AU - Krause, Florian
N1 - Funding information: We would like to thank Julia Volker, Leibniz University Hannover, for valuable technical assistance. Initial screening for crystallization conditions was done by Prof. Dr. George N. Phillips, Jr., University of Wisconsin-Madison, USA, within the Protein Structure Initiative. We gratefully thank the staff scientists at the synchrotron beamline ID23-1, ESRF/Grenoble, for their assistance during diffraction data collection. This work was partially financed by the German Research Foundation (PA 764/10-1). RF was supported by the German Research Foundation (FE 1510/2-1). JWM was funded by the European Commission (Marie Curie Fellowship 625451 SUPA-HD, Sulphation pathways in Health and Disease). The publication of this article was funded by the Open Access Fund of the Leibniz Universität Hannover.
PY - 2017/12/1
Y1 - 2017/12/1
N2 - Sulphotransferases are a diverse group of enzymes catalysing the transfer of a sulfuryl group from 3′-phosphoadenosine 5′-phosphosulphate (PAPS) to a broad range of secondary metabolites. They exist in all kingdoms of life. In Arabidopsis thaliana (L.) Heynh. twenty-two sulphotransferase (SOT) isoforms were identified. Three of those are involved in glucosinolate (Gl) biosynthesis, glycosylated sulphur-containing aldoximes containing chemically different side chains, whose break-down products are involved in stress response against herbivores, pathogens, and abiotic stress. To explain the differences in substrate specificity of desulpho (ds)-Gl SOTs and to understand the reaction mechanism of plant SOTs, we determined the first high-resolution crystal structure of the plant ds-Gl SOT AtSOT18 in complex with 3′-phosphoadenosine 5′-phosphate (PAP) alone and together with the Gl sinigrin. These new structural insights into the determination of substrate specificity were complemented by mutagenesis studies. The structure of AtSOT18 invigorates the similarity between plant and mammalian sulphotransferases, which illustrates the evolutionary conservation of this multifunctional enzyme family. We identified the essential residues for substrate binding and catalysis and demonstrated that the catalytic mechanism is conserved between human and plant enzymes. Our study indicates that the loop-gating mechanism is likely to be a source of the substrate specificity in plants.
AB - Sulphotransferases are a diverse group of enzymes catalysing the transfer of a sulfuryl group from 3′-phosphoadenosine 5′-phosphosulphate (PAPS) to a broad range of secondary metabolites. They exist in all kingdoms of life. In Arabidopsis thaliana (L.) Heynh. twenty-two sulphotransferase (SOT) isoforms were identified. Three of those are involved in glucosinolate (Gl) biosynthesis, glycosylated sulphur-containing aldoximes containing chemically different side chains, whose break-down products are involved in stress response against herbivores, pathogens, and abiotic stress. To explain the differences in substrate specificity of desulpho (ds)-Gl SOTs and to understand the reaction mechanism of plant SOTs, we determined the first high-resolution crystal structure of the plant ds-Gl SOT AtSOT18 in complex with 3′-phosphoadenosine 5′-phosphate (PAP) alone and together with the Gl sinigrin. These new structural insights into the determination of substrate specificity were complemented by mutagenesis studies. The structure of AtSOT18 invigorates the similarity between plant and mammalian sulphotransferases, which illustrates the evolutionary conservation of this multifunctional enzyme family. We identified the essential residues for substrate binding and catalysis and demonstrated that the catalytic mechanism is conserved between human and plant enzymes. Our study indicates that the loop-gating mechanism is likely to be a source of the substrate specificity in plants.
UR - http://www.scopus.com/inward/record.url?scp=85048947930&partnerID=8YFLogxK
U2 - 10.1038/s41598-017-04539-2
DO - 10.1038/s41598-017-04539-2
M3 - Article
C2 - 28646214
AN - SCOPUS:85048947930
VL - 7
JO - Scientific reports
JF - Scientific reports
SN - 2045-2322
IS - 1
M1 - 4160
ER -