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Literature summary for 6.3.4.4 extracted from

  • Mehrotra, S.; B Ningappa, M.; Raman, J.; Anand, R.P.; Balaram, H.
    Mutational analysis of cysteine 328 and cysteine 368 at the interface of Plasmodium falciparum adenylosuccinate synthetase (2012), Biochim. Biophys. Acta, 1824, 589-597.
    View publication on PubMed
Search for more literature for 6.3.4.4

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Plasmodium falciparum

Protein Variants

Protein Variants Comment Organism
C328D the mutant shows reduced Km and increased turnover number for L-aspartate compared to the wild type protein Plasmodium falciparum
C328D/C368D the mutant shows reduced Km and turnover number for L-aspartate compared to the wild type protein Plasmodium falciparum
C328S the mutant exhibits no change in the aspartate Km value but reduced turnover number compared to the wild type protein Plasmodium falciparum
C328S/C368S the mutant shows a 4fold reduced Km for aspartate and reduced turnover number compared to the wild type protein Plasmodium falciparum
C368S the mutant exhibits a 1.7fold increase in the aspartate Km value compared to the wild type protein Plasmodium falciparum

General Stability

General Stability Organism
at 1.5 M urea the enzyme dissociates to form monomers and further increase in urea concentration causes unfolding of this species Plasmodium falciparum

Inhibitors

Inhibitors Comment Organism Structure
Cu2+ complete loss in activity within 4 h at 0.5 mM Plasmodium falciparum
additional information not inhibited Mn2+ Plasmodium falciparum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0152
-
 IMP mutant enzyme C328S, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
0.0168
-
 IMP wild type enzyme, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
0.0172
-
 GTP mutant enzyme C328S/C368S, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
0.0198
-
 IMP mutant enzyme C328S/C368S, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
0.0206
-
 GTP mutant enzyme C368S, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
0.0248
-
 IMP mutant enzyme C368S, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
0.0248
-
 GTP wild type enzyme, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
0.0251
-
 GTP mutant enzyme C328D, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
0.0276
-
 IMP mutant enzyme C328D, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
0.0292
-
 IMP mutant enzyme C368D/C368D, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
0.0358
-
 GTP mutant enzyme C368D/C368D, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
0.044
-
 GTP mutant enzyme C328S, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
0.49
-
 L-Asp mutant enzyme C328S/C368S, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
1.4
-
 L-Asp mutant enzyme C328D, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
1.73
-
 L-Asp mutant enzyme C368D/C368D, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
1.9
-
 L-Asp mutant enzyme C328S, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
1.9
-
 L-Asp wild type enzyme, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
3.3
-
 L-Asp mutant enzyme C368S, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ dependent on Plasmodium falciparum

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
50000
-
 2 * 50000, SDS-PAGE Plasmodium falciparum
89000
-
 gel filtration Plasmodium falciparum
100000
-
 calculated from amino acid sequence Plasmodium falciparum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
GTP + IMP + L-Asp Plasmodium falciparum
-
 GDP + phosphate + adenylosuccinate
-
 ?

Organism

Organism UniProt Comment Textmining
Plasmodium falciparum Q9U8D3
-
-

Purification (Commentary)

Purification (Comment) Organism
ammonium sulfate precipitation, anion exchange chromatography, and gel filtration Plasmodium falciparum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
GTP + IMP + L-Asp
-
 Plasmodium falciparum GDP + phosphate + adenylosuccinate
-
 ?

Subunits

Subunits Comment Organism
homodimer 2 * 50000, SDS-PAGE Plasmodium falciparum

Synonyms

Synonyms Comment Organism
Adenylosuccinate synthetase
-
 Plasmodium falciparum
AdSS
-
 Plasmodium falciparum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.62
-
 IMP mutant enzyme C328S/C368S, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
0.7
-
 IMP mutant enzyme C368D/C368D, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
0.84
-
 IMP mutant enzyme C328S, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
1
-
 IMP mutant enzyme C368S, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
1.1
-
 IMP wild type enzyme, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum
1.7
-
 IMP mutant enzyme C328D, in 30 mM sodium phosphate, pH 7.4 and at 25°C Plasmodium falciparum

Cofactor

Cofactor Comment Organism Structure
GTP
-
 Plasmodium falciparum