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Literature summary for 4.4.1.28 extracted from

  • Tchong, S.I.; Xu, H.; White, R.H.
    L-Cysteine desulfidase: an [4Fe-4S] enzyme isolated from Methanocaldococcus jannaschii that catalyzes the breakdown of L-cysteine into pyruvate, ammonia, and sulfide (2005), Biochemistry, 44, 1659-1670.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
dithiothreitol the air-inactivated enzyme is activated by reaction with Fe2+ and dithiothreitol in the absence of air Methanocaldococcus jannaschii
methyl viologen 4 mM, 40% stimulation Methanocaldococcus jannaschii

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Methanocaldococcus jannaschii

Protein Variants

Protein Variants Comment Organism
C25A mutation reduces the catalytic efficiency and thermal stability of the enzyme Methanocaldococcus jannaschii
C282A mutation reduces the catalytic efficiency and thermal stability of the enzyme Methanocaldococcus jannaschii
C322S mutation reduces the catalytic efficiency and thermal stability of the enzyme Methanocaldococcus jannaschii
C329S mutation reduces the catalytic efficiency and thermal stability of the enzyme Methanocaldococcus jannaschii
C332A/C329A complete loss of activity Methanocaldococcus jannaschii
D323N about 45% loss of specific activity Methanocaldococcus jannaschii
H139N about 10% loss of specific activity Methanocaldococcus jannaschii

Inhibitors

Inhibitors Comment Organism Structure
EDTA 7.1 mM, 70% inhibition Methanocaldococcus jannaschii
hydroxylamine 34.5 mM, 12% inhibition Methanocaldococcus jannaschii
iodoacetamide 0.42 mM, half-life: 12 min. 17 mM, preincubation completely abolishes activity Methanocaldococcus jannaschii
N-ethylmaleimide 0.42 mM, half-life: 8 min Methanocaldococcus jannaschii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.78
-
L-cysteine 65°C, pH 6.6, wild-type enzyme Methanocaldococcus jannaschii

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ the air-inactivated enzyme is activated by reaction with Fe2+ and dithiothreitol in the absence of air Methanocaldococcus jannaschii
Iron-sulfur cluster the air-inactivated enzyme contains 3 mol of iron per subunit Methanocaldococcus jannaschii

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
43000
-
3 * 43000, SDS-PAGE Methanocaldococcus jannaschii
135000
-
gel filtration Methanocaldococcus jannaschii

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-cysteine + H2O Methanocaldococcus jannaschii possible involvement in iron-sulfur center biosynthesis sulfide + NH3 + pyruvate
-
?
L-cysteine + H2O Methanocaldococcus jannaschii DSM 2661 possible involvement in iron-sulfur center biosynthesis sulfide + NH3 + pyruvate
-
?

Organism

Organism UniProt Comment Textmining
Methanocaldococcus jannaschii Q58431
-
-
Methanocaldococcus jannaschii DSM 2661 Q58431
-
-

Oxidation Stability

Oxidation Stability Organism
activity is completely lost by incubating the native protein solution in air for 15 min prior to the incubation Methanocaldococcus jannaschii

Purification (Commentary)

Purification (Comment) Organism
-
Methanocaldococcus jannaschii

Reaction

Reaction Comment Organism Reaction ID
2-aminoprop-2-enoate = 2-iminopropanoate (1b), spontaneous Methanocaldococcus jannaschii
2-iminopropanoate + H2O = pyruvate + NH3 (1c), spontaneous Methanocaldococcus jannaschii
L-cysteine + H2O = sulfide + NH3 + pyruvate overall reaction Methanocaldococcus jannaschii
L-cysteine = 2-aminoprop-2-enoate + sulfide (1a) Methanocaldococcus jannaschii

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.22
-
65°C, pH 6.6, mutant enzyme C25A Methanocaldococcus jannaschii
2.8
-
65°C, pH 6.6, mutant enzyme C282A Methanocaldococcus jannaschii
6.5
-
65°C, pH 6.6, mutant enzyme C322S Methanocaldococcus jannaschii
6.8
-
65°C, pH 6.6, mutant enzyme C329S Methanocaldococcus jannaschii
30
-
65°C, pH 6.6, mutant enzyme D323N Methanocaldococcus jannaschii
49
-
65°C, pH 6.6, mutant enzyme H139N Methanocaldococcus jannaschii
55.8
-
65°C, pH 6.6, wild-type enzyme Methanocaldococcus jannaschii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-cysteine + H2O possible involvement in iron-sulfur center biosynthesis Methanocaldococcus jannaschii sulfide + NH3 + pyruvate
-
?
L-cysteine + H2O the enzyme is very specific for L-cysteine, with no activity being detected with D-cysteine, L-homocysteine, 3-mercaptopropionic acid (cysteine without the amino group), cysteamine (cysteine without the carboxylic acid), or mercaptolactate (the hydroxyl analogue of cysteine) Methanocaldococcus jannaschii sulfide + NH3 + pyruvate
-
?
L-cysteine + H2O possible involvement in iron-sulfur center biosynthesis Methanocaldococcus jannaschii DSM 2661 sulfide + NH3 + pyruvate
-
?
L-cysteine + H2O the enzyme is very specific for L-cysteine, with no activity being detected with D-cysteine, L-homocysteine, 3-mercaptopropionic acid (cysteine without the amino group), cysteamine (cysteine without the carboxylic acid), or mercaptolactate (the hydroxyl analogue of cysteine) Methanocaldococcus jannaschii DSM 2661 sulfide + NH3 + pyruvate
-
?

Subunits

Subunits Comment Organism
trimer 3 * 43000, SDS-PAGE Methanocaldococcus jannaschii

Synonyms

Synonyms Comment Organism
L-cysteine desulfhydrase
-
Methanocaldococcus jannaschii
MJ1025 locus name Methanocaldococcus jannaschii

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
90
-
10 min, 22% loss of activity, mutant enzyme C278A Methanocaldococcus jannaschii
90
-
10 min, 65% loss of activity, mutant enzyme C318S Methanocaldococcus jannaschii
90
-
10 min, 80% loss of activity, mutant enzyme C325S Methanocaldococcus jannaschii
90
-
10 min, no loss of activity, wild-type enzyme Methanocaldococcus jannaschii
100
-
10 min, 80% loss of activity, wild-type enzyme Methanocaldococcus jannaschii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
39.6
-
L-cysteine 65°C, pH 6.6, wild-type enzyme Methanocaldococcus jannaschii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
sharp pH-optiumum Methanocaldococcus jannaschii

pH Range

pH Minimum pH Maximum Comment Organism
6.5 8.5 activities at pH 6.5 and 8.5 are less than 20% of that observed at pH 7.6 Methanocaldococcus jannaschii

Cofactor

Cofactor Comment Organism Structure
additional information both the isolated and recombinant enzymes are devoid of pyridoxal 5'-phosphate Methanocaldococcus jannaschii

General Information

General Information Comment Organism
physiological function possible involvement in iron-sulfur center biosynthesis Methanocaldococcus jannaschii

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
22.2
-
L-cysteine 65°C, pH 6.6, wild-type enzyme Methanocaldococcus jannaschii