Literature summary for 2.5.1.10 extracted from

Maheshwari, S.; Kim, Y.S.; Aripirala, S.; Murphy, M.; Amzel, L.M.; Gabelli, S.B.
Identifying structural determinants of product specificity in Leishmania major farnesyl diphosphate synthase (2020), Biochemistry, 59, 2751-2759 .

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli BL21 (DE3) cells	Leishmania major

Crystallization (Commentary)

Crystallization (Comment)	Organism
crystals of mutant enzymes E97Y, T164Y, and T164W, are produced using the hanging drop vapor diffusion technique at 18°C. Proteins are co-crystallized with inhibitor 476A and substrate IPP in the presence of MgCl	Leishmania major
hanging drop vapor diffusion technique at 18°C. Crystal structures of mutant enzymes E97Y, T164W, and T164Y bound with 3-butyl-1-(2,2-diphosphonoethyl)pyridinium, isopentenyl diphosphate, and modeled farnesyl diphosphate provide strong evidence that these mutations produce the observed effects by altering the size of the binding pocket for the growing isoprenoid chain in the active site of the enzyme	Leishmania major

Crystallization (Comment)

Organism

crystals of mutant enzymes E97Y, T164Y, and T164W, are produced using the hanging drop vapor diffusion technique at 18°C. Proteins are co-crystallized with inhibitor 476A and substrate IPP in the presence of MgCl

Leishmania major

hanging drop vapor diffusion technique at 18°C. Crystal structures of mutant enzymes E97Y, T164W, and T164Y bound with 3-butyl-1-(2,2-diphosphonoethyl)pyridinium, isopentenyl diphosphate, and modeled farnesyl diphosphate provide strong evidence that these mutations produce the observed effects by altering the size of the binding pocket for the growing isoprenoid chain in the active site of the enzyme

Leishmania major

Protein Variants

Protein Variants	Comment	Organism
T164F	mutation completely abolishes the activity	Leishmania major
T164F	mutation completely abolishes the activity of the enzyme. No significant thermal shift between the wild-type enzyme and the mutants, indicating that the single-point mutations do not disrupt the folding or stability of the enzyme	Leishmania major
T164W	mutation completely abolishes the activity	Leishmania major
T164W	mutation completely abolishes the activity of the enzyme. No significant thermal shift between the wild-type enzyme and the mutants, indicating that the single-point mutations do not disrupt the folding or stability of the enzyme	Leishmania major
T164Y	mutation displays dual product specificity and produces a mixture geranyl diphosphate and farnesyl diphosphate as final products, with an activity for farnesyl diphosphate synthesis that is lower than that of the wild-type enzyme	Leishmania major
T164Y	mutation displays dual product specificity and produces a mixture geranyl diphosphate and farnesyl diphosphate as final products, with an activity for farnesyl diphosphate synthesis that is lower than that of the wild-type enzyme. No significant thermal shift between the wild-type enzyme and the mutants, indicating that the single-point mutations do not disrupt the folding or stability of the enzyme	Leishmania major

Inhibitors

Inhibitors	Comment	Organism	Structure
3-butyl-1-(2,2-diphosphonoethyl)pyridinium	-	Leishmania major

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
dimethylallyl diphosphate + isopentenyl diphosphate	Leishmania major	the enzyme catalyzes the sequential condensation of dimethylallyl diphosphate (C5) with isopentenyl diphosphate (IPP) and the resulting geranyl diphosphate (GPP) with another molecule of IPP, eventually producing farnesyl diphosphate which is a precursor for the biosynthesis of a vast majority of isoprenoids	diphosphate + geranyl diphosphate	-	?
dimethylallyl diphosphate + isopentenyl diphosphate	Leishmania major	the enzyme catalyzes the sequential condensation of dimethylallyl diphosphate with isopentenyl diphosphate and the resulting geranyl diphosphate with another molecule of isopentenyl diphosphate, eventually producing farnesyl diphosphate	diphosphate + geranyl diphosphate	-	?
geranyl diphosphate + isopentenyl diphosphate	Leishmania major	the enzyme catalyzes the sequential condensation of dimethylallyl diphosphate (C5) with isopentenyl diphosphate (IPP) and the resulting geranyl diphosphate (GPP) with another molecule of IPP, eventually producing farnesyl diphosphate which is a precursor for the biosynthesis of a vast majority of isoprenoids	diphosphate + (2E,6E)-farnesyl diphosphate	-	?
geranyl diphosphate + isopentenyl diphosphate	Leishmania major	the enzyme catalyzes the sequential condensation of dimethylallyl diphosphate with isopentenyl diphosphate and the resulting geranyl diphosphate with another molecule of isopentenyl diphosphate, eventually producing farnesyl diphosphate	diphosphate + (2E,6E)-farnesyl diphosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Leishmania major	Q4QBL1	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Leishmania major

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
dimethylallyl diphosphate + isopentenyl diphosphate	the enzyme catalyzes the sequential condensation of dimethylallyl diphosphate (C5) with isopentenyl diphosphate (IPP) and the resulting geranyl diphosphate (GPP) with another molecule of IPP, eventually producing farnesyl diphosphate which is a precursor for the biosynthesis of a vast majority of isoprenoids	Leishmania major	diphosphate + geranyl diphosphate	-	?
dimethylallyl diphosphate + isopentenyl diphosphate	the enzyme catalyzes the sequential condensation of dimethylallyl diphosphate with isopentenyl diphosphate and the resulting geranyl diphosphate with another molecule of isopentenyl diphosphate, eventually producing farnesyl diphosphate	Leishmania major	diphosphate + geranyl diphosphate	-	?
dimethylallyl diphosphate + isopentenyl diphosphate	the enzyme catalyzes the sequential condensation of dimethylallyl diphosphate (C5) with isopentenyl diphosphate (IPP) and the resulting geranyl diphosphate (GPP) with another molecule of IPP, eventually producing farnesyl diphosphate	Leishmania major	diphosphate + geranyl diphosphate	-	?
geranyl diphosphate + isopentenyl diphosphate	the enzyme catalyzes the sequential condensation of dimethylallyl diphosphate (C5) with isopentenyl diphosphate (IPP) and the resulting geranyl diphosphate (GPP) with another molecule of IPP, eventually producing farnesyl diphosphate which is a precursor for the biosynthesis of a vast majority of isoprenoids	Leishmania major	diphosphate + (2E,6E)-farnesyl diphosphate	-	?
geranyl diphosphate + isopentenyl diphosphate	the enzyme catalyzes the sequential condensation of dimethylallyl diphosphate with isopentenyl diphosphate and the resulting geranyl diphosphate with another molecule of isopentenyl diphosphate, eventually producing farnesyl diphosphate	Leishmania major	diphosphate + (2E,6E)-farnesyl diphosphate	-	?
geranyl diphosphate + isopentenyl diphosphate	the enzyme catalyzes the sequential condensation of dimethylallyl diphosphate (C5) with isopentenyl diphosphate (IPP) and the resulting geranyl diphosphate (GPP) with another molecule of IPP, eventually producing farnesyl diphosphate	Leishmania major	diphosphate + (2E,6E)-farnesyl diphosphate	-	?

Subunits

Subunits	Comment	Organism
homodimer	-	Leishmania major

Synonyms

Synonyms	Comment	Organism
farnesyl diphosphate synthase	-	Leishmania major
FPPS	-	Leishmania major

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
94	-	Tm-value above, wild-type and mutant enzymes	Leishmania major

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	-	Leishmania major

pH Stability

pH Stability	pH Stability Maximum	Comment	Organism
7.5	8	most stable from pH 7.5 to 8.0	Leishmania major