Literature summary for 4.6.1.19 extracted from

Kimura, K.; Numata, T.; Kakuta, Y.; Kimura, M.
Amino acids conserved at the C-terminal half of the ribonuclease T2 family contribute to protein stability of the enzymes (2004), Biosci. Biotechnol. Biochem., 68, 1748-1757.

Cloned(Commentary)

Cloned (Comment)	Organism
mutants A105L, D107A, P125A, G127A, G144A, V165A and F190A are expressed in Escherichia coli. Mutants Y101A, F102A, L162A and G173A are expressed in Pichia pastoris	Momordica charantia

Crystallization (Commentary)

Crystallization (Comment)	Organism
mutant Y101A in complex with 5'-UMP, resolution at 2.0 A	Momordica charantia

Protein Variants

Protein Variants	Comment	Organism
A105L	significant loss of activity, significant decrease in thermostability	Momordica charantia
D107A	little effect on thermostability	Momordica charantia
F102A	considerable less stable than the wild-type in guanidine-HCl, significant decrease in thermostability	Momordica charantia
F190A	significant decrease in thermostability	Momordica charantia
G127A	moderate decrease in thermostability	Momordica charantia
G144A	moderate decrease in thermostability	Momordica charantia
G173A	little effect on thermostability	Momordica charantia
L162A	unstable	Momordica charantia
P125A	moderate decrease in thermostability	Momordica charantia
V165A	moderate decrease in thermostability	Momordica charantia
Y101A	considerable less stable than the wild-type in guanidine-HCl, significant decrease in thermostability	Momordica charantia

General Stability

General Stability	Organism
the wild-type enzyme remains intact during incubation with chymotrypsin for 8 h (pH 8.0, 37°C). Trypsin causes a time-dependent decrease of the RNase MC1 band concomitant with the increase of one peptide. Mutant Y101A is completely degraded by chymotrypsin and trypsin after 2 h. Mutant F102A is also susceptible to protease degradation. In mutants A105L, D107A, P125A, G127A, G144A, V165A, F190A and G173A the degradation pattern is similar to the wild-type.	Momordica charantia

Organism

the wild-type enzyme remains intact during incubation with chymotrypsin for 8 h (pH 8.0, 37°C). Trypsin causes a time-dependent decrease of the RNase MC1 band concomitant with the increase of one peptide. Mutant Y101A is completely degraded by chymotrypsin and trypsin after 2 h. Mutant F102A is also susceptible to protease degradation. In mutants A105L, D107A, P125A, G127A, G144A, V165A, F190A and G173A the degradation pattern is similar to the wild-type.

Momordica charantia

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
21190	-	monomer, calculated from amino acid sequence	Momordica charantia

Organic Solvent Stability

Organic Solvent	Comment	Organism
guanidine-HCl	mutants Y101A (guanidine-HCl concentration at the midpoint of the transition is 1.59 M) and F102A (guanidine-HCl concentration at the midpoint of the transition is 1.64 M) are considerable less stable than the wild-type (guanidine-HCl concentration at the midpoint of the transition is 2.57 M). In mutant P125A guanidine-HCl concentration at the midpoint of the transition is 2.11 M, in mutant G127A guanidine-HCl concentration at the midpoint of the transition is 2.12 M, in mutant G173A guanidine-HCl concentration at the midpoint of the transition is 2.56 M	Momordica charantia

Organism

Organism	UniProt	Comment	Textmining
Momordica charantia	P23540	bitter gourd	-

Source Tissue

Source Tissue	Comment	Organism	Textmining

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
cytidylyl-3',5'-uridine	37°C, pH 5.5	Momordica charantia	?	-	?

Synonyms

Synonyms	Comment	Organism
ribonuclease MC1	-	Momordica charantia
ribonuclease T2	-	Momordica charantia
RNase MC1	-	Momordica charantia

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
47.2	-	Tm, mutant Y101A	Momordica charantia
50.8	-	Tm, mutant F102A	Momordica charantia
56.1	-	Tm, mutant F190A	Momordica charantia
56.9	-	Tm, mutant A105L	Momordica charantia
58.4	-	Tm, mutant V165A	Momordica charantia
60.5	-	Tm, mutant G144A	Momordica charantia
61.7	-	Tm, mutant G127A	Momordica charantia
62.2	-	Tm, mutant P125A	Momordica charantia
63.6	-	Tm, mutant D107A	Momordica charantia
64.2	-	Tm, mutant G173A	Momordica charantia
64.4	-	Tm, wild-type	Momordica charantia