Literature summary for 2.3.1.97 extracted from

Brannigan, J.A.; Smith, B.A.; Yu, Z.; Brzozowski, A.M.; Hodgkinson, M.R.; Maroof, A.; Price, H.P.; Meier, F.; Leatherbarrow, R.J.; Tate, E.W.; Smith, D.F.; Wilkinson, A.J.
N-Myristoyltransferase from Leishmania donovani: Structural and functional characterisation of a potential drug target for visceral leishmaniasis (2009), J. Mol. Biol., 396, 985-999.

Application

Application	Comment	Organism
drug development	the enzyme is a potential target for development of therapeutic agents against visceral leishmaniasis	Leishmania donovani
drug development	the enzyme is a potential target for drug development	Leishmania donovani

Cloned(Commentary)

Cloned (Comment)	Organism
DNA and amino acid sequence determination and analysis, overexpression of the His-tagged NMT in Escherichia coli strain BL21(DE3) pRareS	Leishmania donovani
overexpression in Escherichia coli strain BL21(DE3)	Leishmania donovani

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme in complex with the non-hydrolysable substrate analogue S-(2-oxo)pentadecyl-CoA, X-ray diffraction structure determination and analysis at 1.4 A resolution, molecular replacement method	Leishmania donovani
purified recombinant His-tagged NMT in complex with the non-hydrolysable substrate analogue S-(2-oxo)pentadecyl-CoA, hanging drop vapour diffusion method, mixing of 0.00125 ml of protein solution containing 8 mg/ml protein in 50 mM Tris, pH 8.0, 50 mM NaCl , with 0.00125 l of reservoir solution containing 0.6 M lithium chloride, 20% w/v PEG 6000, 4% v/v 1,4-butanediol in 0.5 M sodium citrate, pH 4.0, equilibration against 0.8 ml of reservoir solution, 20°C, X-ray diffraction structure determination and analysis at 1.4 A resolution, molecular replacement	Leishmania donovani

Crystallization (Comment)

Organism

purified recombinant enzyme in complex with the non-hydrolysable substrate analogue S-(2-oxo)pentadecyl-CoA, X-ray diffraction structure determination and analysis at 1.4 A resolution, molecular replacement method

Leishmania donovani

purified recombinant His-tagged NMT in complex with the non-hydrolysable substrate analogue S-(2-oxo)pentadecyl-CoA, hanging drop vapour diffusion method, mixing of 0.00125 ml of protein solution containing 8 mg/ml protein in 50 mM Tris, pH 8.0, 50 mM NaCl , with 0.00125 l of reservoir solution containing 0.6 M lithium chloride, 20% w/v PEG 6000, 4% v/v 1,4-butanediol in 0.5 M sodium citrate, pH 4.0, equilibration against 0.8 ml of reservoir solution, 20°C, X-ray diffraction structure determination and analysis at 1.4 A resolution, molecular replacement

Leishmania donovani

Inhibitors

Inhibitors	Comment	Organism	Structure
additional information	ligand binding structure-function analysis for drug design, overview	Leishmania donovani
S-(2-oxo)pentadecyl-CoA	i.e. NHM, a non-hydrolysable myristoyl-CoA analogue, binding structure analysis, overview; i.e. NHM, a non-hydrolysable myristoyl-CoA analogue, binding structure, overview	Leishmania donovani

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.00023	-	myristoyl-CoA	pH 7.4, 37°C, recombinant enzyme	Leishmania donovani
0.0175	-	ARF peptide	pH 7.4, 37°C, recombinant enzyme	Leishmania donovani

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	-	Leishmania donovani

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
tetradecanoyl-CoA + glycylpeptide	Leishmania donovani	NMT catalyses the attachment of the 14C saturated fatty acid, myristate, to the N-terminal glycine residue of a subset of eukaryotic proteins that function in multiple cellular processes, including vesicular protein trafficking and signal transduction	CoA + N-tetradecanoylglycylpeptide	-	?
tetradecanoyl-CoA + glycylpeptide	Leishmania donovani LV9	NMT catalyses the attachment of the 14C saturated fatty acid, myristate, to the N-terminal glycine residue of a subset of eukaryotic proteins that function in multiple cellular processes, including vesicular protein trafficking and signal transduction	CoA + N-tetradecanoylglycylpeptide	-	?

Organism

Organism	UniProt	Comment	Textmining
Leishmania donovani	D0AB09	-	-
Leishmania donovani LV9	D0AB09	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant enzyme from Escherichia coli strain BL21(DE3)	Leishmania donovani
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) pRareS by metal affinity chromatography and gel filtration, removal of the His-tag by cleavage with HRV 3C protease	Leishmania donovani

Source Tissue

Source Tissue	Comment	Organism	Textmining
promastigote	-	Leishmania donovani	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
additional information	the fatty acyl-CoA is largely buried in the N-terminal lobe, its binding leading to the loosening of a flap, which in unliganded NMT structures, occludes the protein substrate binding site in the carboxy-terminal lobe, substrate binding structure, overview	Leishmania donovani	?	-	?
additional information	the fatty acyl-CoA is largely buried in the N-terminal lobe, its binding leading to the loosening of a flap, which in unliganded NMT structures, occludes the protein substrate binding site in the carboxy-terminal lobe, substrate binding structure, overview	Leishmania donovani LV9	?	-	?
myristoyl-CoA + ARF peptide	C-terminally biotinylated ARF peptide, i.e. GLYVSRLFNRLFQKK(Biotin)-NH2, substrate binding structures, overview	Leishmania donovani	CoA + N-myristoyl-ARF peptide	-	?
tetradecanoyl-CoA + GAAPSKIV-NH2	NMT catalyses the attachment of the 14C saturated fatty acid, myristate, to the N-terminal glycine residue of a protein. The fatty acyl-CoA is largely buried in the N-terminal lobe, its binding leading to the loosening of a flap, which in unliganded NMT structures, occludes the protein substrate binding site in the C-terminal lobe, substrate binding structure, overview	Leishmania donovani	CoA + N-tetradecanoyl-GAAPSKIV-NH2	-	?
tetradecanoyl-CoA + GAAPSKIV-NH2	NMT catalyses the attachment of the 14C saturated fatty acid, myristate, to the N-terminal glycine residue of a protein. The fatty acyl-CoA is largely buried in the N-terminal lobe, its binding leading to the loosening of a flap, which in unliganded NMT structures, occludes the protein substrate binding site in the C-terminal lobe, substrate binding structure, overview	Leishmania donovani LV9	CoA + N-tetradecanoyl-GAAPSKIV-NH2	-	?
tetradecanoyl-CoA + GLTISKLFRR	NMT catalyses the attachment of the 14C saturated fatty acid, myristate, to the N-terminal glycine residue of a protein. The fatty acyl-CoA is largely buried in the N-terminal lobe, its binding leading to the loosening of a flap, which in unliganded NMT structures, occludes the protein substrate binding site in the C-terminal lobe, substrate binding structure, overview	Leishmania donovani	CoA + N-tetradecanoyl-GLTISKLFRR	-	?
tetradecanoyl-CoA + GLTISKLFRR	NMT catalyses the attachment of the 14C saturated fatty acid, myristate, to the N-terminal glycine residue of a protein. The fatty acyl-CoA is largely buried in the N-terminal lobe, its binding leading to the loosening of a flap, which in unliganded NMT structures, occludes the protein substrate binding site in the C-terminal lobe, substrate binding structure, overview	Leishmania donovani LV9	CoA + N-tetradecanoyl-GLTISKLFRR	-	?
tetradecanoyl-CoA + glycylpeptide	NMT catalyses the attachment of the 14C saturated fatty acid, myristate, to the N-terminal glycine residue of a subset of eukaryotic proteins that function in multiple cellular processes, including vesicular protein trafficking and signal transduction	Leishmania donovani	CoA + N-tetradecanoylglycylpeptide	-	?
tetradecanoyl-CoA + glycylpeptide	NMT catalyses the attachment of the 14C saturated fatty acid, myristate, to the N-terminal glycine residue of a subset of eukaryotic proteins that function in multiple cellular processes, including vesicular protein trafficking and signal transduction	Leishmania donovani LV9	CoA + N-tetradecanoylglycylpeptide	-	?
tetradecanoyl-CoA + GLYVSRLFNRLFQKK(biotin)	NMT catalyses the attachment of the 14C saturated fatty acid, myristate, to the N-terminal glycine residue of a protein. The fatty acyl-CoA is largely buried in the N-terminal lobe, its binding leading to the loosening of a flap, which in unliganded NMT structures, occludes the protein substrate binding site in the C-terminal lobe, substrate binding structure, overview	Leishmania donovani	CoA + N-tetradecanoyl-GLYVSRLFNRLFQKK(biotin)	-	?
tetradecanoyl-CoA + GLYVSRLFNRLFQKK(biotin)	NMT catalyses the attachment of the 14C saturated fatty acid, myristate, to the N-terminal glycine residue of a protein. The fatty acyl-CoA is largely buried in the N-terminal lobe, its binding leading to the loosening of a flap, which in unliganded NMT structures, occludes the protein substrate binding site in the C-terminal lobe, substrate binding structure, overview	Leishmania donovani LV9	CoA + N-tetradecanoyl-GLYVSRLFNRLFQKK(biotin)	-	?

Synonyms

Synonyms	Comment	Organism
myristoyl-CoA-protein N-myristoyltransferase	-	Leishmania donovani
N-myristoyltransferase	-	Leishmania donovani
NMT	-	Leishmania donovani

Temperature Optimum [°C]

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

pH Optimum

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

IC50 Value

IC50 Value	IC50 Value Maximum	Comment	Organism	Inhibitor	Structure
0.0000687	-	-	Leishmania donovani	S-(2-oxo)pentadecyl-CoA
0.0687	-	pH 7.4, 37°C, recombinant enzyme	Leishmania donovani	S-(2-oxo)pentadecyl-CoA

General Information

General Information	Comment	Organism
physiological function	NMT function is essential for viability in host cells and in insect stages of the pathogenic protozoan parasite	Leishmania donovani
physiological function	NMT catalyses the attachment of the 14-carbon saturated fatty acid, myristate, to the amino-terminal glycine residue of a subset of eukaryotic proteins that function in multiple cellular processes, including vesicular protein trafficking and signal transduction. In these pathways, N-myristoylation facilitates association of substrate proteins with membranes or the hydrophobic domains of other partner peptides. NMT function is essential for viability in all cell types tested to date	Leishmania donovani