Sulfate assimilation in higher plants. Characterization of a stable intermediate in the adenosine 5′-phosphosulfate reductase reaction


Autoria(s): Weber, Markus; Suter, Marianne; Brunold, Christian; Kopriva, Stanislav
Data(s)

01/06/2000

Resumo

The enzyme catalysing the reduction of adenosine 5′-phosphosulfate (AdoPS) to sulfite in higher plants, AdoPS reductase, is considered to be the key enzyme of assimilatory sulfate reduction. In order to address its reaction mechanism, the APR2 isoform of this enzyme from Arabidopsis thaliana was overexpressed in Escherichia coli and purified to homogeneity. Incubation of the enzyme with [35S]AdoPS at 4 °C resulted in radioactive labelling of the protein. Analysis of APR2 tryptic peptides revealed 35SO2–3 bound to Cys248, the only Cys conserved between AdoPS and prokaryotic phosphoadenosine 5′-phosphosulfate reductases. Consistent with this result, radioactivity could be released from the protein by incubation with thiols, inorganic sulfide and sulfite. The intermediate remained stable, however, after incubation with sulfate, oxidized glutathione or AdoPS. Because truncated APR2, missing the thioredoxin-like C-terminal part, could be labelled even at 37 °C, and because this intermediate was more stable than the complete protein, we conclude that the thioredoxin-like domain was required to release the bound SO2–3 from the intermediate. Taken together, these results demonstrate for the first time the binding of 35SO2–3 from [35S]AdoPS to AdoPS reductase and its subsequent release, and thus contribute to our understanding of the molecular mechanism of AdoPS reduction in plants.

Formato

application/pdf

Identificador

http://boris.unibe.ch/81153/1/EurJBiochem_267_3647.pdf

Weber, Markus; Suter, Marianne; Brunold, Christian; Kopriva, Stanislav (2000). Sulfate assimilation in higher plants. Characterization of a stable intermediate in the adenosine 5′-phosphosulfate reductase reaction. European Journal of Biochemistry, 267(12), pp. 3647-3653. Blackwell Science 10.1046/j.1432-1327.2000.01394.x <http://dx.doi.org/10.1046/j.1432-1327.2000.01394.x>

doi:10.7892/boris.81153

info:doi:10.1046/j.1432-1327.2000.01394.x

urn:issn:0014-2956

Idioma(s)

eng

Publicador

Blackwell Science

Relação

http://boris.unibe.ch/81153/

Direitos

info:eu-repo/semantics/restrictedAccess

Fonte

Weber, Markus; Suter, Marianne; Brunold, Christian; Kopriva, Stanislav (2000). Sulfate assimilation in higher plants. Characterization of a stable intermediate in the adenosine 5′-phosphosulfate reductase reaction. European Journal of Biochemistry, 267(12), pp. 3647-3653. Blackwell Science 10.1046/j.1432-1327.2000.01394.x <http://dx.doi.org/10.1046/j.1432-1327.2000.01394.x>

Palavras-Chave #580 Plants (Botany)
Tipo

info:eu-repo/semantics/article

info:eu-repo/semantics/publishedVersion

PeerReviewed