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Literature summary for 2.4.2.44 extracted from

  • Guan, R.; Ho, M.C., Almo, S.C.; Schramm, V.L.
    Methylthioinosine phosphorylase from Pseudomonas aeruginosa. Structure and annotation of a novel enzyme in quorum sensing (2011), Biochemistry, 50, 1247-1254.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
-
Pseudomonas aeruginosa
expressed in Escherichia coli as a His-tagged fusion protein Pseudomonas aeruginosa

Crystallization (Commentary)

Crystallization (Comment) Organism
crystallization of recombinant enzyme (9 mg/ml) in 50 mM HEPES, pH 7.4, 30% polyethylene glycol monomethyl ether 2000, 0.1 M potassium thiocyanate in the presence of 5 mM S-methyl-5'-thioinosine and 5 mM sulfate by sitting-drop vapor diffusion. Crystal structure of the enzyme in complex with hypoxanthine is determined to 2.8 A resolution Pseudomonas aeruginosa
the crystal structure of P. aeruginosa MTIP (PaMTIP) in complex with hypoxanthine is determined to 2.8 A resolution and reveals a three-fold symmetric homotrimer Pseudomonas aeruginosa

Inhibitors

Inhibitors Comment Organism Structure
5'-methylthio-4'-deaza-1'-aza-2'-deoxy-1'-(9-methylene)-immucillin-H
-
Pseudomonas aeruginosa
5'-methylthio-immucillin-H
-
Pseudomonas aeruginosa
5'-phenylthio-immucillin-H
-
Pseudomonas aeruginosa
S-methyl-5'-thioadenosine competitive inhibitor Pseudomonas aeruginosa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0026
-
5'-methylthioinosine pH 7.4, 25┬░C Pseudomonas aeruginosa
0.0026
-
S-methyl-5'-thioinosine pH 7.4, 25┬░C Pseudomonas aeruginosa
0.023
-
adenosine pH 7.4, 25┬░C Pseudomonas aeruginosa
0.023
-
adenine pH 7.4, 25┬░C Pseudomonas aeruginosa
0.09
-
Inosine pH 7.4, 25┬░C Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
S-methyl-5'-thioinosine + phosphate Pseudomonas aeruginosa the enzyme is involved in catabolism of 5'-methylthioadenosine in Pseudomonas aeruginosa. S-Methyl-5'-thioadenosine is converted to S-methyl-5'-thioinosine and hypoxanthine but not to adenine hypoxanthine + S-methyl-5-thio-alpha-D-ribose 1-phosphate
-
?

Organism

Organism UniProt Comment Textmining
Pseudomonas aeruginosa Q9HZK1
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Pseudomonas aeruginosa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5'-methylthioinosine + phosphate
-
Pseudomonas aeruginosa hypoxanthine + 5-methylthio-alpha-D-ribose 1-phosphate
-
?
adenosine + phosphate weak substrate Pseudomonas aeruginosa adenine + alpha-D-ribose 1-phosphate
-
?
adenosine + phosphate
-
Pseudomonas aeruginosa adenine + D-ribose 1-phosphate
-
?
inosine + phosphate the catalytic efficiency (kcat/Km) for S-methyl-5'-thioinosine is 290 times larger than for inosine Pseudomonas aeruginosa hypoxanthine + alpha-D-ribose 1-phosphate
-
?
inosine + phosphate
-
Pseudomonas aeruginosa hypoxanthine + D-ribose 1-phosphate
-
?
additional information S-methyl-5'-thioadenosine is not a substrate Pseudomonas aeruginosa ?
-
?
S-methyl-5'-thioinosine + phosphate the enzyme is involved in catabolism of 5'-methylthioadenosine in Pseudomonas aeruginosa. S-Methyl-5'-thioadenosine is converted to S-methyl-5'-thioinosine and hypoxanthine but not to adenine Pseudomonas aeruginosa hypoxanthine + S-methyl-5-thio-alpha-D-ribose 1-phosphate
-
?
S-methyl-5'-thioinosine + phosphate the methylthio group of S-methyl-5'-thioinosine is important for both substrate binding and catalysis Pseudomonas aeruginosa hypoxanthine + S-methyl-5-thio-alpha-D-ribose 1-phosphate
-
?

Subunits

Subunits Comment Organism
homotrimer crystal structure Pseudomonas aeruginosa
trimer the crystal structure in complex with hypoxanthine, determined to 2.8 A resolution reveals a 3-fold symmetric homotrimer Pseudomonas aeruginosa

Synonyms

Synonyms Comment Organism
methylthioinosine phosphorylase
-
Pseudomonas aeruginosa
MTI phosphorylase
-
Pseudomonas aeruginosa
MTIP
-
Pseudomonas aeruginosa
PA3004
-
Pseudomonas aeruginosa

Temperature Optimum [┬░C]

Temperature Optimum [┬░C] Temperature Optimum Maximum [┬░C] Comment Organism
25
-
assay at Pseudomonas aeruginosa

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0549
-
adenosine pH 7.4, 25┬░C Pseudomonas aeruginosa
0.0549
-
adenine pH 7.4, 25┬░C Pseudomonas aeruginosa
0.57
-
Inosine pH 7.4, 25┬░C Pseudomonas aeruginosa
4.8
-
5'-methylthioinosine pH 7.4, 25┬░C Pseudomonas aeruginosa
4.8
-
S-methyl-5'-thioinosine pH 7.4, 25┬░C Pseudomonas aeruginosa

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
assay at Pseudomonas aeruginosa

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.000000035
-
5'-phenylthio-immucillin-H pH 7.4, 25┬░C Pseudomonas aeruginosa
0.000000076
-
5'-methylthio-immucillin-H pH 7.4, 25┬░C Pseudomonas aeruginosa
0.00000034
-
5'-methylthio-4'-deaza-1'-aza-2'-deoxy-1'-(9-methylene)-immucillin-H pH 7.4, 25┬░C Pseudomonas aeruginosa
0.00000066
-
5'-phenylthio-immucillin-H pH 7.4, 25┬░C Pseudomonas aeruginosa
0.0000008
-
5'-methylthio-4'-deaza-1'-aza-2'-deoxy-1'-(9-methylene)-immucillin-H pH 7.4, 25┬░C Pseudomonas aeruginosa
0.00000084
-
5'-methylthio-immucillin-H pH 7.4, 25┬░C Pseudomonas aeruginosa
0.07
-
S-methyl-5'-thioadenosine pH 7.4, 25┬░C Pseudomonas aeruginosa

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.24
-
adenine pH 7.4, 25┬░C Pseudomonas aeruginosa
6.3
-
Inosine pH 7.4, 25┬░C Pseudomonas aeruginosa
1846
-
S-methyl-5'-thioinosine pH 7.4, 25┬░C Pseudomonas aeruginosa