Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Hartmann, R.; Justesen, J.; Sarkar, S.N.; Sen, G.C.; Yee, V.C.
    Crystal structure of the 2'-specific and double-stranded RNA-activated interferon-induced antiviral protein 2'-5'-oligoadenylate synthetase (2003), Mol. Cell, 12, 1173-1185.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.7.7.84 dsRNA 2'-5'-oligoadenylate synthases are activated by viral double-stranded RNA in infected cells and initiate a cellular response by synthesizing 2'-5'-oligoadenylates, which in turn activate RNase L. All mammalian OAS proteins require dsRNA for activity ssRNA or DNA does not activate this class of enzymes. The dsRNA activator must be at least 15 nucleotides long, and no modification of the 2'-hydroxyl group is tolerated. OAS1 RNA activation site structure, overview Sus scrofa
2.7.7.84 viral dsRNA 2'-5'-oligoadenylate synthases are activated by viral double-stranded RNA in infected cells and initiate a cellular response by synthesizing 2'-5'-oligoadenylates, which in turn activate RNase L. All mammalian OAS proteins require dsRNA for activity ssRNA or DNA does not activate this class of enzymes. The dsRNA activator must be at least 15 nucleotides long, and no modification of the 2'-hydroxyl group is tolerated. OAS1 RNA activation site structure, overview Homo sapiens

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.7.84 OAS1, expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Sus scrofa

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.7.84 OAS1, purified recombinant porcine OAS1, 1.5 mg/ml protein in 50 mM NaCl, 10 mM HEPES, pH 6.8, 0.5 mM EDTA, with 5 mM of iodoacetamide for 30 min at room temperature, then concentrated to 6 mg/ml for crystallization by vapor diffusion, best crystals grow at 20°C in 1:1 drops with a well solution of 30% PEG 2000 MME, 0.2 M ammonium sulfate, and 0.1 M sodium cacodylate at pH 6, X-ray diffraction structure determination and analysis at 2.2 A resolution, molecular replacement Sus scrofa
2.7.7.84 OAS1, X-ray diffraction structure determination and analysis Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
2.7.7.84 K203E site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Sus scrofa
2.7.7.84 K212A the mutant has strongly impaired catalytic activity, KM for ATP is increased by almost 4fold relative to that of the wild-type protein, and kcat is decreased by roughly 8fold Sus scrofa
2.7.7.84 K41E site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Sus scrofa
2.7.7.84 K59E site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Sus scrofa
2.7.7.84 R194E site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Sus scrofa
2.7.7.84 R194E/R198E site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Sus scrofa
2.7.7.84 R198E site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Sus scrofa
2.7.7.84 R198M site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Sus scrofa
2.7.7.84 R245E/K246E site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Sus scrofa
2.7.7.84 R38E site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Sus scrofa
2.7.7.84 R38E/K41E site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Sus scrofa
2.7.7.84 R38E/K41E/K59E/R194E/R198E site-directed mutagenesis, the mutant is almost inactive Sus scrofa
2.7.7.84 S62A the mutant displays a strong substrate inhibition at higher ATP concentrations, more dramatic than that observed for wild-type OAS1 Sus scrofa
2.7.7.84 S63A the mutant displays a strong substrate inhibition at higher ATP concentrations, more dramatic than that observed for wild-type OAS1 Sus scrofa

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.7.84 ATP substrate inhibition at higher concentrations Sus scrofa

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.7.84 0.00044
-
 ATP wild-type OAS1, with ATP as sole substrate, pH and temperature not specified in the publication Sus scrofa
2.7.7.84 0.0011
-
 NAD+ OAS1 mutant S62A, with ATP and NAD+ as substrates, pH and temperature not specified in the publication Sus scrofa
2.7.7.84 0.0013
-
 NAD+ wild-type OAS1, with ATP and NAD+ as substrates, pH and temperature not specified in the publication Sus scrofa
2.7.7.84 0.0014
-
 NAD+ OAS1 mutant S63A, with ATP and NAD+ as substrates, pH and temperature not specified in the publication Sus scrofa
2.7.7.84 0.0017
-
 ATP OAS1 mutant K212A, with ATP as sole substrate, pH and temperature not specified in the publication Sus scrofa

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.7.84 3 ATP Homo sapiens
-
 pppA2'p5'A2'p5'A + 2 diphosphate
-
 ?
2.7.7.84 3 ATP Sus scrofa
-
 pppA2'p5'A2'p5'A + 2 diphosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.84 Homo sapiens
-
 isozyme OAS1
-
2.7.7.84 Sus scrofa
-
-
-
2.7.7.84 Sus scrofa Q29599 isozyme OAS1
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.7.84 recombinant wild-type and mutant OAS1 from Escherichia coli strain BL21(DE3) by gel filtration, ammonium sulfate fractionation, heparin affinity chromatography and gel filtration Sus scrofa

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.7.7.84 0.0005
-
 OAS1 mutant R38E/K41E/K59E/R194E/R198E, pH and temperature not specified in the publication Sus scrofa
2.7.7.84 0.001
-
 OAS1 mutant R194E/R198E, pH and temperature not specified in the publication Sus scrofa
2.7.7.84 0.004
-
 OAS1 mutant R198E, pH and temperature not specified in the publication Sus scrofa
2.7.7.84 0.007
-
 OAS1 mutant R198M, pH and temperature not specified in the publication Sus scrofa
2.7.7.84 0.016
-
 OAS1 mutant R38E/K41E, pH and temperature not specified in the publication Sus scrofa
2.7.7.84 0.05
-
 OAS1 mutant K203E, pH and temperature not specified in the publication Sus scrofa
2.7.7.84 0.117
-
 OAS1 mutant K59E, pH and temperature not specified in the publication Sus scrofa
2.7.7.84 0.16
-
 OAS1 mutant K41E, pH and temperature not specified in the publication Sus scrofa
2.7.7.84 0.17
-
 OAS1 mutant R194E, pH and temperature not specified in the publication Sus scrofa
2.7.7.84 0.22
-
 OAS1 mutant R38E, pH and temperature not specified in the publication Sus scrofa
2.7.7.84 3.3
-
 OAS1 mutant R245E/K246E, pH and temperature not specified in the publication Sus scrofa
2.7.7.84 10.1
-
 wild-type OAS1, pH and temperature not specified in the publication Sus scrofa

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.84 3 ATP
-
 Homo sapiens pppA2'p5'A2'p5'A + 2 diphosphate
-
 ?
2.7.7.84 3 ATP
-
 Sus scrofa pppA2'p5'A2'p5'A + 2 diphosphate
-
 ?
2.7.7.84 dATP + NAD+
-
 Sus scrofa ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.7.7.84 2'-5'-oligoadenylate synthetase
-
 Homo sapiens
2.7.7.84 2'-5'-oligoadenylate synthetase
-
 Sus scrofa
2.7.7.84 OAS
-
 Homo sapiens
2.7.7.84 OAS
-
 Sus scrofa

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.7.84 0.95
-
 ATP OAS1 mutant K212A, with ATP as sole substrate, pH and temperature not specified in the publication Sus scrofa
2.7.7.84 0.99
-
 NAD+ OAS1 mutant S62A, with ATP and NAD+ as substrates, pH and temperature not specified in the publication Sus scrofa
2.7.7.84 3.6
-
 NAD+ OAS1 mutant S63A, with ATP and NAD+ as substrates, pH and temperature not specified in the publication Sus scrofa
2.7.7.84 3.6
-
 NAD+ wild-type OAS1, with ATP and NAD+ as substrates, pH and temperature not specified in the publication Sus scrofa
2.7.7.84 8.1
-
 ATP wild-type OAS1, with ATP as sole substrate, pH and temperature not specified in the publication Sus scrofa

Expression

EC Number Organism Comment Expression
2.7.7.84 Homo sapiens induction by interferon up
2.7.7.84 Sus scrofa induction by interferon up

General Information

EC Number General Information Comment Organism
2.7.7.84 evolution conserved catalytic mechanism for the 2'- and 3'-specific nucleotidyl transferases. specific nucleotidyl transferases. Comparison with structures of other superfamily members indicates that the donor substrates are bound by conserved active site features while the acceptor substrates are oriented by nonconserved regions Homo sapiens
2.7.7.84 evolution conserved catalytic mechanism for the 2'- and 3'-specific nucleotidyl transferases. specific nucleotidyl transferases. Comparison with structures of other superfamily members indicates that the donor substrates are bound by conserved active site features while the acceptor substrates are oriented by nonconserved regions Sus scrofa
2.7.7.84 malfunction mutation of the conserved Leu3 and Pro7 and of Cys330, Cys331, and Lys332 reduce enzyme activity Homo sapiens
2.7.7.84 malfunction mutation of the conserved Leu3 and Pro7 and of Cys330, Cys331, and Lys332 reduce enzyme activity. Mutants S62A and S63A display Michaelis-Menten kinetics toward NAD+, the kcat of the Ser62Ala mutant is approximately 3fold lower than the kcat for either the wild-type or the S63A mutant Sus scrofa
2.7.7.84 additional information OAS active site structure with three conserved active site aspartic acid residues, overview Homo sapiens
2.7.7.84 additional information OAS active site structure with three conserved active site aspartic acid residues, overview Sus scrofa
2.7.7.84 physiological function 2'-5'-oligoadenylate synthases are interferon-induced, double-stranded RNA-activated antiviral enzymes which are the only proteins known to catalyze 2'-specific nucleotidyl transfer Homo sapiens
2.7.7.84 physiological function 2'-5'-oligoadenylate synthases are interferon-induced, double-stranded RNA-activated antiviral enzymes which are the only proteins known to catalyze 2'-specific nucleotidyl transfer Sus scrofa