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Literature summary for 2.1.1.20 extracted from

  • Luka, Z.; Wagner, C.
    Expression and purification of glycine N-methyltransferases in Escherichia coli (2003), Protein Expr. Purif., 28, 280-286.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
when expressed in pTYB vector as a fusion protein with intein and the chitin binding domain, a cleavage of intein is found. The cleavage takes place at two sites near the N-terminus of intein and results in the appearance of an abnormal GNMT protein after one-column cleavage of the fusion protein, which can not be separated from normal GNMT. For this reason expression is done in the vector pET-17b. Expression of soluble protein in Escherichia coli at about 20-40 mg/L Mus musculus
when expressed in pTYB vector as a fusion protein with intein and the chitin binding domain, a cleavage of intein is found. The cleavage takes place at two sites near the N-terminus of intein and results in the appearance of an abnormal GNMT protein after one-column cleavage of the fusion protein, which can not be separated from normal GNMT. For this reason expression is done in the vector pET-17b. Expression of soluble protein in Escherichia coli at about 20-40 mg/L Homo sapiens
when expressed in pTYB vector as a fusion protein with intein and the chitin binding domain, a cleavage of intein is found. The cleavage takes place at two sites near the N-terminus of intein and results in the appearance of an abnormal GNMT protein after one-column cleavage of the fusion protein, which can not be separated from normal GNMT. For this reason expression is done in the vector pET-17b. Expression of soluble protein in Escherichia coli at about 20-40 mg/L Rattus norvegicus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
130000
-
gel filtration Mus musculus
130000
-
gel filtration Homo sapiens
130000
-
gel filtration Rattus norvegicus

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-
Mus musculus
-
-
-
Rattus norvegicus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Mus musculus
-
Homo sapiens
-
Rattus norvegicus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
Homo sapiens

Storage Stability

Storage Stability Organism
4°C, 5.0 mM DTT, 2.0 mM EDTA, 0.02% NaN3, stable for at least 2 weeks Mus musculus
4°C, 5.0 mM DTT, 2.0 mM EDTA, 0.02% NaN3, stable for at least 2 weeks Homo sapiens
4°C, 5.0 mM DTT, 2.0 mM EDTA, 0.02% NaN3, stable for at least 2 weeks Rattus norvegicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
S-adenosyl-L-methionine + glycine mechanism: the bound S-adenosyl-L-methionine is firmly connected to protein and a „Gly pocket" is created near the bound S-adenosyl-L-methionine. The second substrate Gly binds to Arg175 and is brought into the Gly pocket. Five hydrogen bonds connect the Gly in the proximity of the bound S-adenosyl-L-methionine and orient the lone pair orbital on the amino nitrogen of Gly towards the donor methyl group of S-adenosyl-L-methionine. Thermal motion of the enzyme leads to a collision of the N and C(E) so that a SN2 methyltransfer reaction occurs Mus musculus S-adenosyl-L-homocysteine + sarcosine
-
?
S-adenosyl-L-methionine + glycine mechanism: the bound S-adenosyl-L-methionine is firmly connected to protein and a „Gly pocket" is created near the bound S-adenosyl-L-methionine. The second substrate Gly binds to Arg175 and is brought into the Gly pocket. Five hydrogen bonds connect the Gly in the proximity of the bound S-adenosyl-L-methionine and orient the lone pair orbital on the amino nitrogen of Gly towards the donor methyl group of S-adenosyl-L-methionine. Thermal motion of the enzyme leads to a collision of the N and C(E) so that a SN2 methyltransfer reaction occurs Homo sapiens S-adenosyl-L-homocysteine + sarcosine
-
?
S-adenosyl-L-methionine + glycine mechanism: the bound S-adenosyl-L-methionine is firmly connected to protein and a „Gly pocket" is created near the bound S-adenosyl-L-methionine. The second substrate Gly binds to Arg175 and is brought into the Gly pocket. Five hydrogen bonds connect the Gly in the proximity of the bound S-adenosyl-L-methionine and orient the lone pair orbital on the amino nitrogen of Gly towards the donor methyl group of S-adenosyl-L-methionine. Thermal motion of the enzyme leads to a collision of the N and C(E) so that a SN2 methyltransfer reaction occurs Rattus norvegicus S-adenosyl-L-homocysteine + sarcosine
-
?

Subunits

Subunits Comment Organism
tetramer
-
Mus musculus
tetramer
-
Homo sapiens
tetramer
-
Rattus norvegicus

Synonyms

Synonyms Comment Organism
GNMT
-
Mus musculus
GNMT
-
Homo sapiens
GNMT
-
Rattus norvegicus