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Literature summary for 2.1.2.1 extracted from

  • Chang, W.N.; Tsai, J.N.; Chen, B.H.; Huang, H.S.; Fu, T.F.
    Serine hydroxymethyltransferase isoforms are differentially inhibited by leucovorin: characterization and comparison of recombinant zebrafish serine hydroxymethyltransferases (2007), Drug Metab. Dispos., 35, 2127-2137.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Danio rerio

Inhibitors

Inhibitors Comment Organism Structure
leucovorin leucovorin (N5-CHO-THF) exhibits a differential inhibition pattern: it significantly inhibits the aldol cleavage of serine catalyzed by zebrafish cytosolic SHMT but it inhibits to a lesser extent the reaction catalyzed by the mitochondrial isozyme. Approximately 70% and 30% inhibition are observed for zebrafish cytosolic- and zebrafish mitochondrial SHMT activities, respectively, in the presence of 70 mM leucovorin. An even larger difference between both isoenzymes is observed when the inhibition is assayed in the presence of 50 mM serine; leucovorin (N5-CHO-THF) exhibits a differential inhibition pattern: it significantly inhibits the aldol cleavage of serine catalyzed by zebrafish cytosolic SHMT but it inhibits to a lesser extent the reaction catalyzed by the mitochondrial isozyme. Approximately 70% and 30% inhibition are observed for zebrafish cytosolic- and zebrafish mitochondrial SHMT activities, respectively, in the presence of 70 mM leucovorin. An even larger difference between both isoenzymes is observed when the inhibition is assayed in the presence of 50 mM serine Danio rerio

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.43
-
serine
-
Danio rerio

Localization

Localization Comment Organism GeneOntology No. Textmining
cytosol zcSHMT is detected in the cytosol as well as in the nucleus Danio rerio 5829
-
mitochondrion peptide sequence alignment with the known SHMT from other species and the prospective zebrafish cytosolic SHMT as well as signal prediction software reveals a potential mitochondrial signal peptide cleavage site between residues 20 and 30 Danio rerio 5739
-
nucleus zcSHMT is detected in the cytosol as well as in the nucleus Danio rerio 5634
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
50000
-
SDS-PAGE Danio rerio

Organism

Organism UniProt Comment Textmining
Danio rerio A9LDD9
-
-
Danio rerio Q7SXN1
-
-

Purification (Commentary)

Purification (Comment) Organism
using a 30 to 50% ammonium sulfate precipitation, gel filtration and chromatography on CM-Sephadex columns. From 2 liters of cells, approximately 43 mg of pure zmSHMT is obtained with an overall yield of 90% Danio rerio

Source Tissue

Source Tissue Comment Organism Textmining
heart elevated mRNA expression detected by RT-PCT Danio rerio
-
liver elevated mRNA expression detected by RT-PCT. Strong protein expression detected by western blotting Danio rerio
-
liver siginificant protein expression detected by Western Blotting only in liver and ovary Danio rerio
-
additional information RT-PCR results show that zmSHMT mRNA is evenly distributed among tissues. However zmSHMT protein expression does not correspond to the detected mRNA expression detected in most tissues Danio rerio
-
ovary elevated mRNA expression detected by RT-PCT. Strong protein expression detected by western blotting Danio rerio
-
ovary siginificant protein expression detected by Western Blotting only in liver and ovary Danio rerio
-

Storage Stability

Storage Stability Organism
-20°C (or -80°C), in the presence of 10% glycerol, for at least 6 months, no significant change in catalytic activity Danio rerio

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5,10-methylenetetrahydrofolate + glycine + H2O
-
Danio rerio tetrahydrofolate + L-serine
-
r

Subunits

Subunits Comment Organism
tetramer 4 * 50000, gel filtration Danio rerio

Synonyms

Synonyms Comment Organism
zcSHMT
-
Danio rerio
zebrafish cytosolic serine hydroxymethyltransferase
-
Danio rerio
zebrafish mitochondiral serine hydroxymethyltransferase
-
Danio rerio
zmSHMT
-
Danio rerio

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Danio rerio

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
8.95
-
serine
-
Danio rerio

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Danio rerio

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate
-
Danio rerio

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
30
-
-
Danio rerio leucovorin
70
-
-
Danio rerio leucovorin