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Literature summary for 2.1.1.41 extracted from

  • Ganapathy, K.; Jones, C.W.; Stephens, C.M.; Vatsyayan, R.; Marshall, J.A.; Nes, W.D.
    Molecular probing of the Saccharomyces cerevisiae sterol 24-C methyltransferase reveals multiple amino acid residues involved with C2-transfer activity (2008), Biochim. Biophys. Acta, 1781, 344-351.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Saccharomyces cerevisiae
expressed in Escherichia coli, recombinant protein, site-directed mutagenesis Saccharomyces cerevisiae

Protein Variants

Protein Variants Comment Organism
A193L no activity Saccharomyces cerevisiae
A193L no enzyme activity Saccharomyces cerevisiae
A193S 95% relative enzyme activity Saccharomyces cerevisiae
A193S relative activity: 95% Saccharomyces cerevisiae
A196L 73% relative enzyme activity Saccharomyces cerevisiae
A196L relative activity: 73% Saccharomyces cerevisiae
A200L 4% relative enzyme activity Saccharomyces cerevisiae
A200L relative activity: 4% Saccharomyces cerevisiae
A221L 25% relative enzyme activity Saccharomyces cerevisiae
A221L relative activity: 25% Saccharomyces cerevisiae
A350L 100% relative enzyme activity Saccharomyces cerevisiae
A350L relative activity: 100% Saccharomyces cerevisiae
C128L no activity Saccharomyces cerevisiae
C128L no enzyme activity Saccharomyces cerevisiae
C198L 95% relative enzyme activity Saccharomyces cerevisiae
C198L relative activity: 95% Saccharomyces cerevisiae
C198V 16% relative enzyme activity Saccharomyces cerevisiae
C198V relative activity: 16% Saccharomyces cerevisiae
D125L no activity Saccharomyces cerevisiae
D152L no activity Saccharomyces cerevisiae
D189L relative activity: 91% Saccharomyces cerevisiae
D229L relative activity: 100% Saccharomyces cerevisiae
D276L no activity Saccharomyces cerevisiae
D65L relative activity: 100% Saccharomyces cerevisiae
D79L relative activity: 81% Saccharomyces cerevisiae
E108L relative activity: 34% Saccharomyces cerevisiae
E195L no activity Saccharomyces cerevisiae
E209L relative activity: 79% Saccharomyces cerevisiae
E224L relative activity: 74% Saccharomyces cerevisiae
E246L relative activity: 6% Saccharomyces cerevisiae
E64L relative activity: 100% Saccharomyces cerevisiae
E68L no activity Saccharomyces cerevisiae
E82D 55% relative enzyme activity Saccharomyces cerevisiae
E82D relative activity: 55% Saccharomyces cerevisiae
E82L relative activity: 99% Saccharomyces cerevisiae
E82Q 90% relative enzyme activity Saccharomyces cerevisiae
E82Q relative activity: 90% Saccharomyces cerevisiae
E98L relative activity: 100% Saccharomyces cerevisiae
F178L 34% relative enzyme activity Saccharomyces cerevisiae
F178L relative activity: 34% Saccharomyces cerevisiae
F183L 100% relative enzyme activity Saccharomyces cerevisiae
F188L 2% relative enzyme activity Saccharomyces cerevisiae
F188L relative activity: 2% Saccharomyces cerevisiae
F220L 29% relative enzyme activity Saccharomyces cerevisiae
F220L relative activity: 29% Saccharomyces cerevisiae
F269L no activity Saccharomyces cerevisiae
F269L no enzyme activity Saccharomyces cerevisiae
F357L 100% relative enzyme activity Saccharomyces cerevisiae
F357L relative activity: 100% Saccharomyces cerevisiae
F89L 100% relative enzyme activity Saccharomyces cerevisiae
F89L relative activity: 100% Saccharomyces cerevisiae
F91L 15% relative enzyme activity Saccharomyces cerevisiae
F91L relative activity: 15% Saccharomyces cerevisiae
G127L 1% relative enzyme activity Saccharomyces cerevisiae
G127L relative activity: 1% Saccharomyces cerevisiae
G129L no activity Saccharomyces cerevisiae
G129L no enzyme activity Saccharomyces cerevisiae
G131L 5% relative enzyme activity Saccharomyces cerevisiae
G131L relative activity: 5% Saccharomyces cerevisiae
G132L 50% relative enzyme activity Saccharomyces cerevisiae
G132L relative activity: 50% Saccharomyces cerevisiae
G217L 100% relative enzyme activity, mutant can perform C2-transfer activity to give 24-ethyl(idene)-sterols Saccharomyces cerevisiae
G217L relative activity: 100% Saccharomyces cerevisiae
G218L 100% relative enzyme activity, mutant can perform C2-transfer activity to give 24-ethyl(idene)-sterols Saccharomyces cerevisiae
G218L relative activity: 100% Saccharomyces cerevisiae
G347L 5% relative enzyme activity Saccharomyces cerevisiae
G347L relative activity: 5% Saccharomyces cerevisiae
G351L 90% relative enzyme activity Saccharomyces cerevisiae
G351L relative activity: 90% Saccharomyces cerevisiae
G352L 3% relative enzyme activity Saccharomyces cerevisiae
G352L relative activity: 3% Saccharomyces cerevisiae
G84L 14% relative enzyme activity Saccharomyces cerevisiae
G84L relative activity: 14% Saccharomyces cerevisiae
G86L 16% relative enzyme activity Saccharomyces cerevisiae
G86L relative activity: 16% Saccharomyces cerevisiae
H107L relative activity: 23% Saccharomyces cerevisiae
H199L relative activity: 2% Saccharomyces cerevisiae
H199Q 45% relative enzyme activity Saccharomyces cerevisiae
H199Q relative activity: 45% Saccharomyces cerevisiae
H199R 20% relative enzyme activity Saccharomyces cerevisiae
H199R relative activity: 20% Saccharomyces cerevisiae
H238L relative activity: 85% Saccharomyces cerevisiae
H90L no activity Saccharomyces cerevisiae
H90Q no activity Saccharomyces cerevisiae
H90Q no enzyme activity Saccharomyces cerevisiae
H90R no activity Saccharomyces cerevisiae
H90R no enzyme activity Saccharomyces cerevisiae
I194L 67% relative enzyme activity Saccharomyces cerevisiae
I194L relative activity: 67% Saccharomyces cerevisiae
K215L 100% relative enzyme activity Saccharomyces cerevisiae
K215L relative activity: 100% Saccharomyces cerevisiae
K353L 56% relative enzyme activity Saccharomyces cerevisiae
additional information scanning mutagenesis experiments involving a leucine replacement of 52 amino acids in Erg6p followed by substitution of key residues with functionally or structurally similar amino acids indicate that 5 new residues at positions Y192, G217, G218, T219 and Y223 can switch the course of C1-transfer activity to include plant-like C2-transfer activity. The data support a model in which several conserved and non-conserved amino acids located in distinct regions of the Saccharomyces cerevisiae Erg6p regulate the course of the C-methylation reaction toward product differences Saccharomyces cerevisiae
P133L no activity Saccharomyces cerevisiae
P133L no enzyme activity Saccharomyces cerevisiae
P201L 58% relative enzyme activity Saccharomyces cerevisiae
P201L relative activity: 58% Saccharomyces cerevisiae
P216L 65% relative enzyme activity Saccharomyces cerevisiae
P216L relative activity: 65% Saccharomyces cerevisiae
S354L 100% relative enzyme activity Saccharomyces cerevisiae
S354L relative activity: 100% Saccharomyces cerevisiae
S87E 100% relative enzyme activity Saccharomyces cerevisiae
S87L 100% relative enzyme activity Saccharomyces cerevisiae
S87L relative activity: 100% Saccharomyces cerevisiae
S87Q 100% relative enzyme activity Saccharomyces cerevisiae
S88L 61% relative enzyme activity Saccharomyces cerevisiae
S88L relative activity: 61% Saccharomyces cerevisiae
S88N 17% relative enzyme activity Saccharomyces cerevisiae
S88N relative activity: 17% Saccharomyces cerevisiae
S97E relative activity: 100% Saccharomyces cerevisiae
S97Q relative activity: 100% Saccharomyces cerevisiae
T197L 93% relative enzyme activity Saccharomyces cerevisiae
T197L relative activity: 93% Saccharomyces cerevisiae
T219L 32% relative enzyme activity, mutant can perform C2-transfer activity to give 24-ethyl(idene)-sterols Saccharomyces cerevisiae
V126L 1% relative enzyme activity Saccharomyces cerevisiae
V126L relative activity: 1% Saccharomyces cerevisiae
V130L 100% relative enzyme activity Saccharomyces cerevisiae
V130L relative activity: 100% Saccharomyces cerevisiae
V222L 27% relative enzyme activity Saccharomyces cerevisiae
V349L 47% relative enzyme activity Saccharomyces cerevisiae
V349L relative activity: 47% Saccharomyces cerevisiae
W225F 100% relative enzyme activity Saccharomyces cerevisiae
W225F relative activity: 60% Saccharomyces cerevisiae
W225L 100% relative enzyme activity Saccharomyces cerevisiae
W225L relative activity: 100% Saccharomyces cerevisiae
W286F 100% relative enzyme activity Saccharomyces cerevisiae
W286F relative activity: 51% Saccharomyces cerevisiae
W286L 2% relative enzyme activity Saccharomyces cerevisiae
W286L relative activity: 2% Saccharomyces cerevisiae
W85F 100% relative enzyme activity Saccharomyces cerevisiae
W85F relative activity: 55% Saccharomyces cerevisiae
W85L 40% relative enzyme activity Saccharomyces cerevisiae
W85L relative activity: 40% Saccharomyces cerevisiae
Y153F 100% relative enzyme activity Saccharomyces cerevisiae
Y153F relative activity: 41% Saccharomyces cerevisiae
Y153L no enzyme activity Saccharomyces cerevisiae
Y153L relative activity: 100% Saccharomyces cerevisiae
Y192F 100% relative enzyme activity, mutant can perform C2-transfer activity to give 24-ethyl(idene)-sterols Saccharomyces cerevisiae
Y192L 95% relative enzyme activity Saccharomyces cerevisiae
Y207F 100% relative enzyme activity Saccharomyces cerevisiae
Y207F relative activity: 100% Saccharomyces cerevisiae
Y207L 3% relative enzyme activity Saccharomyces cerevisiae
Y207L relative activity: 3% Saccharomyces cerevisiae
Y223F 100% relative enzyme activity, mutant can perform C2-transfer activity to give 24-ethyl(idene)-sterols Saccharomyces cerevisiae
Y223F relative activity: 100% Saccharomyces cerevisiae
Y223L 100% relative enzyme activity Saccharomyces cerevisiae
Y223L relative activity: 100% Saccharomyces cerevisiae
Y287L 20% relative enzyme activity Saccharomyces cerevisiae
Y287L relative activity: 20% Saccharomyces cerevisiae
Y74F 100% relative enzyme activity Saccharomyces cerevisiae
Y74F relative activity: 100% Saccharomyces cerevisiae
Y74L 100% relative enzyme activity Saccharomyces cerevisiae
Y74L relative activity: 100% Saccharomyces cerevisiae
Y81F relative activity: 100% Saccharomyces cerevisiae
Y81L 90% relative enzyme activity Saccharomyces cerevisiae
Y81L relative activity: 90% Saccharomyces cerevisiae
Y83F 95% relative enzyme activity Saccharomyces cerevisiae
Y83F relative activity: 95% Saccharomyces cerevisiae
Y83L 90% relative enzyme activity Saccharomyces cerevisiae
Y83L relative activity: 90% Saccharomyces cerevisiae
Y83W 100% relative enzyme activity Saccharomyces cerevisiae
Y83W relative activity: 100% Saccharomyces cerevisiae

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
Saccharomyces cerevisiae

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae
-
-
-
Saccharomyces cerevisiae P25087
-
-

Purification (Commentary)

Purification (Comment) Organism
gel filtration Saccharomyces cerevisiae

Source Tissue

Source Tissue Comment Organism Textmining
commercial preparation
-
Saccharomyces cerevisiae
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
prediction of secondary structure, eight residues at positions 79, 81, 82, 192, 217, 218, 219 and 223 critical for catalysis, scanning mutagenesis experiments involving a leucine replacement of 52 amino acids, substitution of key residues with functionally or structurally similar amino acids, 5 new residues at positions Y192, G217, G218, T219 and Y223 can switch the course of C1-transfer activity to include plant-like C2-transfer activity, model supported in which several conserved and non-conserved amino acids located in distinct regions regulate course of the C-methylation reaction toward product differences Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information formation of the ergostane (C1-transfer activity) and stigmastane (C2-transfer activity) sterol side chains, stepwise mutagenesis of amino acids of Erg6p, identification of five residues within the putative active site that contribute directly to C-methylation pathways operated by fungal and plant SMT proteins Saccharomyces cerevisiae ?
-
?
S-adenosyl-L-methionine + zymosterol
-
Saccharomyces cerevisiae S-adenosyl-L-homocysteine + fecosterol
-
?

Synonyms

Synonyms Comment Organism
Erg6p
-
Saccharomyces cerevisiae
SMT protein
-
Saccharomyces cerevisiae
SMT1
-
Saccharomyces cerevisiae
sterol C24-methyltransferase
-
Saccharomyces cerevisiae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
35
-
assay at Saccharomyces cerevisiae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Saccharomyces cerevisiae