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Literature summary for 2.5.1.21 extracted from

  • Bhargava, P.; Kumar, K.; Chaudhaery, S.S.; Saxena, A.K.; Roy, U.
    Cloning, overexpression and characterization of Leishmania donovani squalene synthase (2010), FEMS Microbiol. Lett., 311, 82-92.
    View publication on PubMed

Application

Application Comment Organism
drug development LdSSN is a potential drug target for visceral leishmaniasis Leishmania donovani

Cloned(Commentary)

Cloned (Comment) Organism
LdSSN DNA and amino acid sequence determination and analysis, sequence comparison, phylogenetic analysis, expression as C-terminally His6-tagged protein in Escherichia coli strain BL21 (DE3) Leishmania donovani

Inhibitors

Inhibitors Comment Organism Structure
Guanidinium chloride loss of 86% activity at 0.3M GdmCl, 50% at 0.2 M Leishmania donovani
Urea loss of 81% activity at 2 M, 50% at 1 M Leishmania donovani
zaragozic acid A is a potent inhibitor of mammalian SSN and also a competitive inhibitor of recombinant Leishmania donovani SSN, 50% inhibition at 100 nM Leishmania donovani

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics, recombinant LdSSN, overview Leishmania donovani
0.0038
-
(2E,6E)-farnesyl diphosphate pH 7.4, 37°C Leishmania donovani
0.04323
-
NADPH pH 7.4, 37°C Leishmania donovani

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Leishmania donovani

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2 (2E,6E)-farnesyl diphosphate + NADPH + H+ Leishmania donovani overall reaction squalene + 2 diphosphate + NADP+
-
?
2 (2E,6E)-farnesyl diphosphate + NADPH + H+ Leishmania donovani MHOM/IN/80/Dd8 overall reaction squalene + 2 diphosphate + NADP+
-
?

Organism

Organism UniProt Comment Textmining
Leishmania donovani Q257D4
-
-
Leishmania donovani MHOM/IN/80/Dd8 Q257D4
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged LdSSN from Escherichia coli strain BL21(DE3) by anion exchange chromatography, nickel affinity chrmatography is not successful Leishmania donovani

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2 (2E,6E)-farnesyl diphosphate + NADPH + H+ overall reaction Leishmania donovani squalene + 2 diphosphate + NADP+
-
?
2 (2E,6E)-farnesyl diphosphate + NADPH + H+ overall reaction Leishmania donovani MHOM/IN/80/Dd8 squalene + 2 diphosphate + NADP+
-
?

Subunits

Subunits Comment Organism
More sequence comparison, overview Leishmania donovani

Synonyms

Synonyms Comment Organism
SSN
-
Leishmania donovani

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
-
Leishmania donovani

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 45 83% and 88% activity at a temperature of 30°C and 45°C Leishmania donovani

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
60
-
purified recombinant enzyme, pH 7.4, loss of 85% activity Leishmania donovani

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
-
Leishmania donovani

pH Range

pH Minimum pH Maximum Comment Organism
6 9 activity range, over 90% of maximal activity at pH 7.0-8.0, 30% at pH 6.0, 20% at pH 9.0 Leishmania donovani

Cofactor

Cofactor Comment Organism Structure
NAD(P)H
-
Leishmania donovani

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.000074
-
zaragozic acid A pH 7.4, 37°C Leishmania donovani

General Information

General Information Comment Organism
metabolism squalene synthase is a major enzyme in the sterol biosynthetic pathway Leishmania donovani
physiological function squalene synthase catalyses an unusual head-to-head reductive dimerization of two molecules of farnesyl-pyrophosphate in a two-step reaction to form squalene Leishmania donovani