Details
| Original language | English |
|---|---|
| Pages (from-to) | 1291-1304 |
| Number of pages | 14 |
| Journal | FEBS Journal |
| Volume | 272 |
| Issue number | 5 |
| Publication status | Published - 1 Mar 2005 |
Abstract
In several organisms D-cysteine desulfhydrase (D-CDes) activity (EC 4.1.99.4) was measured; this enzyme decomposes D-cysteine into pyruvate, H 2S, and NH3. A gene encoding a putative D-CDes protein was identified in Arabidopsis thaliana (L) Heynh. based on high homology to an Escherichia coli protein called YedO that has D-CDes activity. The deduced Arabidopsis protein consists of 401 amino acids and has a molecular mass of 43.9 kDa. It contains a pyridoxal-5′-phosphate binding site. The purified recombinant mature protein had a Km for D-cysteine of 0.25 mM. Only D-cysteine but not L-cysteine was converted by D-CDes to pyruvate, H 2S, and NH3. The activity was inhibited by aminooxy acetic acid and hydroxylamine, inhibitors specific for pyridoxal-5′-phosphate dependent proteins, at low micromolar concentrations. The protein did not exhibit 1-aminocyclopropane-1-carboxylate deaminase activity (EC 3.5.99.7) as homologous bacterial proteins. Western blot analysis of isolated organelles and localization studies using fusion constructs with the green fluorescent protein indicated an intracellular localization of the nuclear encoded D-CDes protein in the mitochondria. D-CDes RNA levels increased with proceeding development of Arabidopsis but decreased in senescent plants; D-CDes protein levels remained almost unchanged in the same plants whereas specific D-CDes activity was highest in senescent plants. In plants grown in a 12-h light/12-h dark rhythm D-CDes RNA levels were highest in the dark, whereas protein levels and enzyme activity were lower in the dark period than in the light indicating post-translational regulation. Plants grown under low sulfate concentration showed an accumulation of D-CDes RNA and increased protein levels, the D-CDes activity was almost unchanged. Putative in vivo functions of the Arabidopsis D-CDes protein are discussed.
Keywords
- 1-aminocyclopropane-1-carboxylate deaminase, Arabidopsis thaliana, D-cysteine, Desulfhydrase, YedO
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Biology
- Biochemistry, Genetics and Molecular Biology(all)
- Cell Biology
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In: FEBS Journal, Vol. 272, No. 5, 01.03.2005, p. 1291-1304.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Isolation and characterization of a D-cysteine desulfhydrase protein from Arabidopsis thaliana
AU - Riemenschneider, Anja
AU - Wegele, Rosalina
AU - Schmidt, Ahlert
AU - Papenbrock, Jutta
PY - 2005/3/1
Y1 - 2005/3/1
N2 - In several organisms D-cysteine desulfhydrase (D-CDes) activity (EC 4.1.99.4) was measured; this enzyme decomposes D-cysteine into pyruvate, H 2S, and NH3. A gene encoding a putative D-CDes protein was identified in Arabidopsis thaliana (L) Heynh. based on high homology to an Escherichia coli protein called YedO that has D-CDes activity. The deduced Arabidopsis protein consists of 401 amino acids and has a molecular mass of 43.9 kDa. It contains a pyridoxal-5′-phosphate binding site. The purified recombinant mature protein had a Km for D-cysteine of 0.25 mM. Only D-cysteine but not L-cysteine was converted by D-CDes to pyruvate, H 2S, and NH3. The activity was inhibited by aminooxy acetic acid and hydroxylamine, inhibitors specific for pyridoxal-5′-phosphate dependent proteins, at low micromolar concentrations. The protein did not exhibit 1-aminocyclopropane-1-carboxylate deaminase activity (EC 3.5.99.7) as homologous bacterial proteins. Western blot analysis of isolated organelles and localization studies using fusion constructs with the green fluorescent protein indicated an intracellular localization of the nuclear encoded D-CDes protein in the mitochondria. D-CDes RNA levels increased with proceeding development of Arabidopsis but decreased in senescent plants; D-CDes protein levels remained almost unchanged in the same plants whereas specific D-CDes activity was highest in senescent plants. In plants grown in a 12-h light/12-h dark rhythm D-CDes RNA levels were highest in the dark, whereas protein levels and enzyme activity were lower in the dark period than in the light indicating post-translational regulation. Plants grown under low sulfate concentration showed an accumulation of D-CDes RNA and increased protein levels, the D-CDes activity was almost unchanged. Putative in vivo functions of the Arabidopsis D-CDes protein are discussed.
AB - In several organisms D-cysteine desulfhydrase (D-CDes) activity (EC 4.1.99.4) was measured; this enzyme decomposes D-cysteine into pyruvate, H 2S, and NH3. A gene encoding a putative D-CDes protein was identified in Arabidopsis thaliana (L) Heynh. based on high homology to an Escherichia coli protein called YedO that has D-CDes activity. The deduced Arabidopsis protein consists of 401 amino acids and has a molecular mass of 43.9 kDa. It contains a pyridoxal-5′-phosphate binding site. The purified recombinant mature protein had a Km for D-cysteine of 0.25 mM. Only D-cysteine but not L-cysteine was converted by D-CDes to pyruvate, H 2S, and NH3. The activity was inhibited by aminooxy acetic acid and hydroxylamine, inhibitors specific for pyridoxal-5′-phosphate dependent proteins, at low micromolar concentrations. The protein did not exhibit 1-aminocyclopropane-1-carboxylate deaminase activity (EC 3.5.99.7) as homologous bacterial proteins. Western blot analysis of isolated organelles and localization studies using fusion constructs with the green fluorescent protein indicated an intracellular localization of the nuclear encoded D-CDes protein in the mitochondria. D-CDes RNA levels increased with proceeding development of Arabidopsis but decreased in senescent plants; D-CDes protein levels remained almost unchanged in the same plants whereas specific D-CDes activity was highest in senescent plants. In plants grown in a 12-h light/12-h dark rhythm D-CDes RNA levels were highest in the dark, whereas protein levels and enzyme activity were lower in the dark period than in the light indicating post-translational regulation. Plants grown under low sulfate concentration showed an accumulation of D-CDes RNA and increased protein levels, the D-CDes activity was almost unchanged. Putative in vivo functions of the Arabidopsis D-CDes protein are discussed.
KW - 1-aminocyclopropane-1-carboxylate deaminase
KW - Arabidopsis thaliana
KW - D-cysteine
KW - Desulfhydrase, YedO
UR - http://www.scopus.com/inward/record.url?scp=14644418532&partnerID=8YFLogxK
U2 - 10.1111/j.1742-4658.2005.04567.x
DO - 10.1111/j.1742-4658.2005.04567.x
M3 - Article
C2 - 15720402
AN - SCOPUS:14644418532
VL - 272
SP - 1291
EP - 1304
JO - FEBS Journal
JF - FEBS Journal
SN - 1742-464X
IS - 5
ER -