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Literature summary for 2.1.4.1 extracted from

  • Muenchhoff, J.; Siddiqui, K.S.; Poljak, A.; Raftery, M.J.; Barrow, K.D.; Neilan, B.A.
    A novel prokaryotic L-arginine:glycine amidinotransferase is involved in cylindrospermopsin biosynthesis (2010), FEBS J., 277, 3844-3860.
    View publication on PubMed
Search for more literature for 2.1.4.1

Cloned(Commentary)

Cloned (Comment) Organism
gene cyrA, DNA and amino acid sequence determination and analysis, phylogenetic analysis, overexpression of N-terminal His6-tagged CyrA in Escherichia coli strain BL21(DE3) Cylindrospermopsis raciborskii

Inhibitors

Inhibitors Comment Organism Structure
L-ornithine partial mixed product inhibition, L-ornithine therefore binds to the active site of CyrA at a binding site distinct from the hydroxylamine-binding site Cylindrospermopsis raciborskii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information Michaelis-Menten kinetics with non-natural substrate hydroxylamine, kinetic mechanism of CyrA as a hybrid of ping-pong and sequential mechanisms, overview Cylindrospermopsis raciborskii
3.5
-
 L-arginine pH 8.5, 30°C Cylindrospermopsis raciborskii
6.9
-
 glycine pH 8.5, 30°C Cylindrospermopsis raciborskii

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
45680
-
 2 * 45680, native enzyme, sequence calculation, 2 * 50120, His6-tagged enzyme, sequence calculation Cylindrospermopsis raciborskii
45680
-
 4 * 45680, native enzyme, sequence calculation, 4 * 50120, His6-tagged enzyme, sequence calculation Cylindrospermopsis raciborskii
50120
-
 2 * 45680, native enzyme, sequence calculation, 2 * 50120, His6-tagged enzyme, sequence calculation Cylindrospermopsis raciborskii
50120
-
 4 * 45680, native enzyme, sequence calculation, 4 * 50120, His6-tagged enzyme, sequence calculation Cylindrospermopsis raciborskii
98000
-
 dimeric CyrA, gel filtration Cylindrospermopsis raciborskii
185000
-
 tetrameric CyrA, gel filtration Cylindrospermopsis raciborskii

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-arginine + glycine Cylindrospermopsis raciborskii
-
 L-ornithine + guanidinoacetate
-
 ?
L-arginine + glycine Cylindrospermopsis raciborskii AWT205
-
 L-ornithine + guanidinoacetate
-
 ?

Organism

Organism UniProt Comment Textmining
Cylindrospermopsis raciborskii B0LI36 gene cyrA
-
Cylindrospermopsis raciborskii AWT205 B0LI36 gene cyrA
-

Purification (Commentary)

Purification (Comment) Organism
recombinant N-terminal His6-tagged CyrA from Escherichia coli strain BL21(DE3) Cylindrospermopsis raciborskii

Reaction

Reaction Comment Organism Reaction ID
L-arginine + glycine = L-ornithine + guanidinoacetate reaction and kinetic mechanisms, overview Cylindrospermopsis raciborskii
a

Storage Stability

Storage Stability Organism
4°C, purified CyrA, total loss of activity occurs within 48 h despite addition of reducing agents such as dithiothreitol, Tris(2-carboxyethyl) phosphine, or beta-mercaptoethanol Cylindrospermopsis raciborskii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
hydroxylamine + glycine
-
 Cylindrospermopsis raciborskii L-ornithine + hydroxyguanidine
-
 ?
hydroxylamine + glycine
-
 Cylindrospermopsis raciborskii AWT205 L-ornithine + hydroxyguanidine
-
 ?
L-arginine + glycine
-
 Cylindrospermopsis raciborskii L-ornithine + guanidinoacetate
-
 ?
L-arginine + glycine L-ornithine is formed in the presence of only a single substrate, arginine, and its production therefore does not require the presence of the second substrate, glycine Cylindrospermopsis raciborskii L-ornithine + guanidinoacetate
-
 ?
L-arginine + glycine
-
 Cylindrospermopsis raciborskii AWT205 L-ornithine + guanidinoacetate
-
 ?
L-arginine + glycine L-ornithine is formed in the presence of only a single substrate, arginine, and its production therefore does not require the presence of the second substrate, glycine Cylindrospermopsis raciborskii AWT205 L-ornithine + guanidinoacetate
-
 ?
additional information narrow substrate specificity, no activity with L-homoarginine, agmatine, L-canavanine, guanidine hydrochloride, urea, gamma-guanidinobutyric acid, and beta-guanidinoproprionic acid as amidino group donors, or L-alanine, beta-alanine, gamma-aminobutyric acid, ethanolamine, taurine, L-lysine, alpha-amino-oxyacetic acid and L-norvaline as amidino group acceptors, overview Cylindrospermopsis raciborskii ?
-
 ?
additional information narrow substrate specificity, no activity with L-homoarginine, agmatine, L-canavanine, guanidine hydrochloride, urea, gamma-guanidinobutyric acid, and beta-guanidinoproprionic acid as amidino group donors, or L-alanine, beta-alanine, gamma-aminobutyric acid, ethanolamine, taurine, L-lysine, alpha-amino-oxyacetic acid and L-norvaline as amidino group acceptors, overview Cylindrospermopsis raciborskii AWT205 ?
-
 ?

Subunits

Subunits Comment Organism
dimer 2 * 45680, native enzyme, sequence calculation, 2 * 50120, His6-tagged enzyme, sequence calculation Cylindrospermopsis raciborskii
tetramer 4 * 45680, native enzyme, sequence calculation, 4 * 50120, His6-tagged enzyme, sequence calculation Cylindrospermopsis raciborskii

Synonyms

Synonyms Comment Organism
CyrA
-
 Cylindrospermopsis raciborskii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
32
-
-
 Cylindrospermopsis raciborskii

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
32 40 20% of maximal activity at 40°C, maximal activity at 32°C Cylindrospermopsis raciborskii

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
 inactive CyrA lacks a well-defined tertiary structure at any temperature Cylindrospermopsis raciborskii
44.5
-
 an inactivated form of this enzyme retains an appreciable amount of secondary structure elements even on heating to 94°C, but loses its tertiary structure at low temperature of 44.5°C resulting in a state reminiscent of a molten globule. The presence of 500 mm NaCl decreases Tmax at pH 7.5 from 54°C to 49°C Cylindrospermopsis raciborskii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
875
-
 glycine per active site, pH 8.5, 30°C Cylindrospermopsis raciborskii
875
-
 L-arginine per active site, pH 8.5, 30°C Cylindrospermopsis raciborskii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
-
 Cylindrospermopsis raciborskii

pH Range

pH Minimum pH Maximum Comment Organism
7 8.5 25% of maximal activity at pH 7.0, maximal activity at pH 8.5 Cylindrospermopsis raciborskii

pH Stability

pH Stability pH Stability Maximum Comment Organism
6.5
-
 purified recombinant enzyme, maximally stable at Cylindrospermopsis raciborskii

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.25
-
 L-ornithine pH 8.5, 30°C Cylindrospermopsis raciborskii

pI Value

Organism Comment pI Value Maximum pI Value
Cylindrospermopsis raciborskii sequence calculation, native enzyme
-
 5.1
Cylindrospermopsis raciborskii sequence calculation, His6-tagged enzyme
-
 5.6

General Information

General Information Comment Organism
evolution CyrA is phylogenetically distinct from other amidinotransferases Cylindrospermopsis raciborskii
metabolism CyrA is involved in the pathway for biosynthesis of the polyketide-derived hepatotoxin cylindrospermopsin Cylindrospermopsis raciborskii