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Literature summary for 4.6.1.19 extracted from

  • Suzuki, A.; Yao, M.; Tanaka, I.; Numata, T.; Kikukawa, S.; Yamasaki, N.; Kimura, M.
    Crystal structures of the ribonuclease MC1 from bitter gourd seeds, complexed with 2'-UMP or 3'-UMP, reveal structural basis for uridine specificity (2000), Biochem. Biophys. Res. Commun., 275, 572-576.
    View publication on PubMed
Search for more literature for 4.6.1.19

Cloned(Commentary)

Cloned (Comment) Organism
expression of mutant enzymes in Escherichia coli BL21(DE3) Momordica charantia

Crystallization (Commentary)

Crystallization (Comment) Organism
19 mg/ml wild-type enzyme in complex with 2'-UMP and 3'-UMP, hanging drop vapour diffusion method, reservoir solution contains 0.2 M sodium acetate, 0.1 M sodium cacodylate, pH 6.7, 21% PEG 8000, 10 mM 2'-UMP or 3'-UMP, 4 days, X-ray diffraction structure determination and analysis at 1.48 and 1.77 A resolution, respectively Momordica charantia

Protein Variants

Protein Variants Comment Organism
L73A site-directed mutagenesis, slightly increased catalytic efficiency and 14.5fold decreased binding affinity compared to the wild-type enzyme Momordica charantia
N71T site-directed mutagenesis, 2.3fold decreased catalytic efficiency and 7.0fold decreased binding affinity compared to the wild-type enzyme Momordica charantia
R74S site-directed mutagenesis, decreased catalytic efficiency and 0.8fold increased binding affinity compared to the wild-type enzyme Momordica charantia
V72L site-directed mutagenesis, catalytic efficiency and binding affinity similar to the wild-type enzyme Momordica charantia

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1
-
 CpU mutant R74S, pH 6.5, 37°C Momordica charantia
1.38
-
 CpU wild-type enzyme, pH 6.5, 37°C Momordica charantia
1.57
-
 CpU mutant V72L, pH 6.5, 37°C Momordica charantia
9.63
-
 CpU mutant N71T, pH 6.5, 37°C Momordica charantia
20
-
 CpU mutant L73A, pH 6.5, 37°C Momordica charantia

Organism

Organism UniProt Comment Textmining
Momordica charantia
-
 bitter gourd
-

Reaction

Reaction Comment Organism Reaction ID
RNA + H2O = an [RNA fragment]-3'-nucleoside-3'-phosphate + a 5'-hydroxy-ribonucleotide-3'-[RNA fragment] substrate recognition, binding and specificity, Asn71 and Leu73 are involved Momordica charantia
a

Source Tissue

Source Tissue Comment Organism Textmining
seed
-
 Momordica charantia
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
CpU + H2O
-
 Momordica charantia ?
-
 ?
additional information enzyme preferentially cleaves the 5'-side of uridine, Asn71 and Leu73 are involved in substrate specificity Momordica charantia ?
-
 ?
RNA + H2O
-
 Momordica charantia ?
-
 ?

Synonyms

Synonyms Comment Organism
More enzyme belongs to the ribonuclease T2 family Momordica charantia
ribonuclease MC1
-
 Momordica charantia
RNase MC1
-
 Momordica charantia

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Momordica charantia

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.37
-
 CpU mutant N71T, pH 6.5, 37°C Momordica charantia
7.38
-
 CpU mutant R74S, pH 6.5, 37°C Momordica charantia
12.3
-
 CpU mutant V72L, pH 6.5, 37°C Momordica charantia
12.42
-
 CpU wild-type enzyme, pH 6.5, 37°C Momordica charantia
13.1
-
 CpU mutant L73A, pH 6.5, 37°C Momordica charantia

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
 assay at Momordica charantia