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Literature summary for 2.1.1.56 extracted from

  • Hausmann, S.; Zheng, S.; Fabrega, C.; Schneller, S.W.; Lima, C.D.; Shuman, S.
    Encephalitozoon cuniculi mRNA cap (guanine N-7) methyltransferase: methyl acceptor specificity, inhibition BY S-adenosylmethionine analogs, and structure-guided mutational analysis (2005), J. Biol. Chem., 280, 20404-20412.
    View publication on PubMed
Search for more literature for 2.1.1.56

Cloned(Commentary)

Cloned (Comment) Organism
inserted into vector pET16b and transformed into Escherichia coli BL21(DE3), yeast strain YBS40 transformed with CEN TRP1 plasmids containing the wild-type and mutant alleles of ECM1 Encephalitozoon cuniculi

Crystallization (Commentary)

Crystallization (Comment) Organism
Ecm1-aza-S-adenosyl-L-methionine binary complex, by vapor diffusion, reveals that the inhibitor occupies the same site as S-adenosyl-L-methionine Encephalitozoon cuniculi

Protein Variants

Protein Variants Comment Organism
D70A lethal, suppresses methyltransferase activity to less than 3% of the wild-type level, in vivo lethality correlates with loss of catalytic activity Encephalitozoon cuniculi
D70E growth at all temperatures Encephalitozoon cuniculi
D70N lethal Encephalitozoon cuniculi
D78A lethal, suppresses methyltransferase activity to less than 3% of the wild-type level, in vivo lethality correlates with loss of catalytic activity Encephalitozoon cuniculi
D78E growth at all temperatures except at 37°C Encephalitozoon cuniculi
D78N growth at all temperatures Encephalitozoon cuniculi
D94A suppresses methyltransferase activity to less than 3% of the wild-type level, in vivo lethality correlates with loss of catalytic activity Encephalitozoon cuniculi
D94E growth at all temperatures Encephalitozoon cuniculi
D94N lethal Encephalitozoon cuniculi
E225A growth at all temperatures except at 37°C Encephalitozoon cuniculi
E225D growth at all temperatures Encephalitozoon cuniculi
E225Q growth at all temperatures Encephalitozoon cuniculi
F141A suppresses methyltransferase activity to less than 3% of the wild-type level, in vivo lethality correlates with loss of catalytic activity Encephalitozoon cuniculi
F214A growth at all temperatures except at 37°C Encephalitozoon cuniculi
F214L growth at all temperatures, slow growth at 37°C Encephalitozoon cuniculi
H144A growth at all temperatures Encephalitozoon cuniculi
K267A growth at all temperatures Encephalitozoon cuniculi
K54A lethal, suppresses methyltransferase activity to less than 3% of the wild-type level, in vivo lethality correlates with loss of catalytic activity Encephalitozoon cuniculi
K54Q lethal Encephalitozoon cuniculi
K54R lethal Encephalitozoon cuniculi
K75Q slow growth at all temperatures Encephalitozoon cuniculi
K75R growth at all temperatures Encephalitozoon cuniculi
K81Q lethal Encephalitozoon cuniculi
K81R lethal Encephalitozoon cuniculi
additional information residues Lys-54, Asp-70, Asp-78, and Asp-94 that comprise the S-adenosyl-L-methionine-binding pocket are all essential for Ecm1 function in yeast, i.e. alanine substitutions at each of these positions are lethal, loss of function caused by the elimination of Phe-141 attests to the importance of its van der Waals interactions with the guanosine methyl acceptor seen in the Ecm1-GTP cocrystal Encephalitozoon cuniculi
N51D lethal Encephalitozoon cuniculi
R106A lethal Encephalitozoon cuniculi
R106K lethal Encephalitozoon cuniculi
R106Q lethal Encephalitozoon cuniculi
R59A growth at all temperatures Encephalitozoon cuniculi
R84A growth at all temperatures Encephalitozoon cuniculi
Y124F growth at all temperatures, slow growth at 37°C Encephalitozoon cuniculi
Y145A lethal Encephalitozoon cuniculi
Y145F growth at all temperatures Encephalitozoon cuniculi
Y145L growth at all temperatures Encephalitozoon cuniculi
Y145S lethal Encephalitozoon cuniculi
Y212A growth at all temperatures except at 37°C Encephalitozoon cuniculi
Y212F growth at all temperatures Encephalitozoon cuniculi

Inhibitors

Inhibitors Comment Organism Structure
aza-S-adenosyl-L-methionine
-
 Encephalitozoon cuniculi
carbocyclic aza-S-adenosyl-L-methionine
-
 Encephalitozoon cuniculi
S-adenosyl-L-homocysteine inhibits methylation of GTP in the presence of 0.005 mM S-adenosyl-L-methionine Encephalitozoon cuniculi
sinefungin
-
 Encephalitozoon cuniculi

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.1
-
 GpppA
-
 Encephalitozoon cuniculi
1
-
 GTP
-
 Encephalitozoon cuniculi
2.4
-
 GDP
-
 Encephalitozoon cuniculi

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
40000
-
 1 * 40000, SDS-PAGE Encephalitozoon cuniculi

Organism

Organism UniProt Comment Textmining
Encephalitozoon cuniculi Q8SR66
-
-

Purification (Commentary)

Purification (Comment) Organism
by nickel-affinity chromatography Encephalitozoon cuniculi

Storage Stability

Storage Stability Organism
-80 °C, Tris-HCl buffer, pH 7.5, 10% glycerol Encephalitozoon cuniculi

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information does not methylate ATP, CTP, UTP, ITP, or m7GTP Encephalitozoon cuniculi ?
-
 ?
S-adenosyl-L-methionine + dGTP
-
 Encephalitozoon cuniculi S-adenosyl-L-homocysteine + m7dGTP
-
 ?
S-adenosyl-L-methionine + GDP
-
 Encephalitozoon cuniculi S-adenosyl-L-homocysteine + m7GDP
-
 ?
S-adenosyl-L-methionine + GpppA
-
 Encephalitozoon cuniculi S-adenosyl-L-homocysteine + m7GpppA
-
 ?
S-adenosyl-L-methionine + GTP
-
 Encephalitozoon cuniculi S-adenosyl-L-homocysteine + m7GTP
-
 ?

Subunits

Subunits Comment Organism
monomer 1 * 40000, SDS-PAGE Encephalitozoon cuniculi

Synonyms

Synonyms Comment Organism
Ecm1
-
 Encephalitozoon cuniculi
mRNA cap (guanine N-7) methyltransferase
-
 Encephalitozoon cuniculi

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
 Encephalitozoon cuniculi

pH Range

pH Minimum pH Maximum Comment Organism
6 7.5 methyl transfer by Ecm1 declines sharply below pH 6.0 Encephalitozoon cuniculi

Cofactor

Cofactor Comment Organism Structure
additional information reaction is cofactor-independent Encephalitozoon cuniculi

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.0015
-
-
 Encephalitozoon cuniculi sinefungin
0.004
-
-
 Encephalitozoon cuniculi S-adenosyl-L-homocysteine
0.035
-
-
 Encephalitozoon cuniculi carbocyclic aza-S-adenosyl-L-methionine
0.1
-
-
 Encephalitozoon cuniculi aza-S-adenosyl-L-methionine