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

BRENDA support

Literature summary extracted from

  • Martin, G.; Moglich, A.; Keller, W.; Doublie, S.
    Biochemical and structural insights into substrate binding and catalytic mechanism of mammalian poly(A) polymerase (2004), J. Mol. Biol., 341, 911-925.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.7.19 expression in Escherichia coli Bos taurus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.7.19 Crystals of PAP complexed to 3'-dATP and Mn2+ are grown from a solution made of 22% (w/v) polyethylene glycol 8000, 100 mM Mes (pH 6), 120 mM ammonium sulfate, 5 mM CaCl2, 2 mM MnCl2, and 2 mM beta-mercaptoethanol. Cocrystallization of PAP with 3'-dATP and MgCl2 is performed under conditions similar to those reported, with the exception that 5 mM MgCl2 is used instead of 2 mM MnCl2. Crystals are of the space group p212121, with one monomer per asymmetric unit and a solvent content of ~55%. Bos taurus

Protein Variants

EC Number Protein Variants Comment Organism
2.7.7.19 D167N strong reduction of catalytic efficiency Bos taurus
2.7.7.19 D167N/N202A sharp decrease in catalytic efficiency Bos taurus
2.7.7.19 D167N/T317G sharp decrease in catalytic efficiency Bos taurus
2.7.7.19 D167N/V247R sharp decrease in catalytic efficiency Bos taurus
2.7.7.19 F100D strong reduction of catalytic efficiency Bos taurus
2.7.7.19 G203H strong reduction of catalytic efficiency Bos taurus
2.7.7.19 K228A increase in Km for ATP Bos taurus
2.7.7.19 K232A increase in Km for ATP Bos taurus
2.7.7.19 N202A high Km value for RNA, strong reduction of catalytic efficiency, incorporates etheno-ATP and N1-methyl-ATP better than wild-type, but incorporates N6-methyl-ATP poorly Bos taurus
2.7.7.19 N239A kcat for ATP is reduced fourfold Bos taurus
2.7.7.19 R199A reduces kcat for RNA about 30-fold Bos taurus
2.7.7.19 T317G increase in Km for ATP, tolerates etheno-ATP and GTP, rejects N6-methyl-ATP Bos taurus
2.7.7.19 V156A strong reduction of catalytic efficiency Bos taurus
2.7.7.19 V247R kcat for ATP is reduced 2000-fold, strong reduction of catalytic efficiency Bos taurus
2.7.7.19 Y237A increase in Km for ATP Bos taurus

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.7.19 CaCl2 remaining activity 3% Bos taurus
2.7.7.19 CoCl2 remaining activity 75% Bos taurus
2.7.7.19 CuCl2 remaining activity 0.1% Bos taurus
2.7.7.19 Mn2+ high concentrations Bos taurus
2.7.7.19 Zn2+ inhibits PAP activity at concentrations above 10 µM in the presence of 5 mM MgCl2 Bos taurus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.7.19 0.136
-
ATP mutant D167N Bos taurus
2.7.7.19 0.154
-
ATP mutant D167N/N202A Bos taurus
2.7.7.19 0.207
-
ATP mutant N239A Bos taurus
2.7.7.19 0.213
-
ATP mutant F153A Bos taurus
2.7.7.19 0.229
-
ATP wild-type, reactions are started with the addition of PAP and incubated at 37 °C for 15 minutes. Reactions are stopped by the addition of 8 microL of stop buffer (formamide with trace amount of bromophenol blue dye) Bos taurus
2.7.7.19 0.232
-
ATP mutant V156A Bos taurus
2.7.7.19 0.247
-
ATP mutant A319R Bos taurus
2.7.7.19 0.248
-
ATP mutant K158A Bos taurus
2.7.7.19 0.255
-
ATP mutant R199A Bos taurus
2.7.7.19 0.256
-
ATP mutant V154A Bos taurus
2.7.7.19 0.266
-
ATP mutant V247A Bos taurus
2.7.7.19 0.275
-
ATP mutant N202A Bos taurus
2.7.7.19 0.297
-
ATP mutant V247R Bos taurus
2.7.7.19 0.298
-
ATP mutant Q323A Bos taurus
2.7.7.19 0.357
-
ATP mutant V206A Bos taurus
2.7.7.19 0.361
-
ATP mutant F100D Bos taurus
2.7.7.19 0.503
-
ATP mutant K232A Bos taurus
2.7.7.19 0.51
-
RNA (A)15 mutant A319R Bos taurus
2.7.7.19 0.627
-
ATP mutant D167N/T317G Bos taurus
2.7.7.19 0.74
-
RNA (A)15 mutant K232A Bos taurus
2.7.7.19 1.002
-
ATP mutant T317G Bos taurus
2.7.7.19 1.34
-
RNA (A)15 wild-type Bos taurus
2.7.7.19 1.402
-
ATP mutant Y237A Bos taurus
2.7.7.19 1.43
-
RNA (A)15 mutant K228A Bos taurus
2.7.7.19 1.47
-
RNA (A)15 mutant V154A Bos taurus
2.7.7.19 1.88
-
RNA (A)15 mutant Q323A Bos taurus
2.7.7.19 2.191
-
ATP mutant K228A Bos taurus
2.7.7.19 2.63
-
RNA (A)15 mutant Y237A Bos taurus
2.7.7.19 2.8
-
RNA (A)15 mutant V206A Bos taurus
2.7.7.19 3.13
-
RNA (A)15 mutant T317G Bos taurus
2.7.7.19 5.31
-
RNA (A)15 mutant K158A Bos taurus
2.7.7.19 6.94
-
RNA (A)15 mutant N239A Bos taurus
2.7.7.19 7.32
-
RNA (A)15 mutant V156A Bos taurus
2.7.7.19 9.85
-
RNA (A)15 mutant V247A Bos taurus
2.7.7.19 10.21
-
RNA (A)15 mutant R199A Bos taurus
2.7.7.19 11.16
-
RNA (A)15 mutant V247R Bos taurus
2.7.7.19 11.44
-
RNA (A)15 mutant N202A Bos taurus
2.7.7.19 12.72
-
RNA (A)15 mutant G203H Bos taurus
2.7.7.19 13.15
-
RNA (A)15 mutant D167N Bos taurus
2.7.7.19 14.37
-
RNA (A)15 mutant F100D Bos taurus
2.7.7.19 16.41
-
RNA (A)15 mutant D167N/N202A Bos taurus
2.7.7.19 18.82
-
RNA (A)15 mutant D167N/T317G Bos taurus
2.7.7.19 22.84
-
RNA (A)15 mutant D167N/V247R Bos taurus
2.7.7.19 24.99
-
RNA (A)15 mutant F153A Bos taurus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.7.19 Mg2+ optimal concentration is about 5 mM Bos taurus

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.19 Bos taurus
-
bovine PAP
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.7.19
-
Bos taurus

Storage Stability

EC Number Storage Stability Organism
2.7.7.19 -80°C, 10-20 mg protein/ml, flash frozen Bos taurus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.19 ATP + RNA (A)15
-
Bos taurus diphosphate + RNA (A)16
-
?
2.7.7.19 ATP + RNA(A)15
-
Bos taurus diphosphate + RNA(A)16
-
?
2.7.7.19 ATP + RNAn adenine can bind in two different configurations in the PAP active site Bos taurus diphosphate + RNAn+1
-
?

Synonyms

EC Number Synonyms Comment Organism
2.7.7.19 PAP
-
Bos taurus
2.7.7.19 poly(A) polymerase
-
Bos taurus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.7.19 0.002
-
ATP mutant V247R Bos taurus
2.7.7.19 0.0025
-
RNA(A)15 mutant D167N/V247R Bos taurus
2.7.7.19 0.006
-
ATP mutant D167N/N202A Bos taurus
2.7.7.19 0.006
-
RNA(A)15 mutant G203H Bos taurus
2.7.7.19 0.007
-
ATP mutant D167N/T317G Bos taurus
2.7.7.19 0.009
-
ATP mutant D167N Bos taurus
2.7.7.19 0.015
-
ATP mutant F100D Bos taurus
2.7.7.19 0.018
-
ATP mutant V156A Bos taurus
2.7.7.19 0.022
-
ATP mutant N202A Bos taurus
2.7.7.19 0.08
-
RNA(A)15 mutant N202A Bos taurus
2.7.7.19 0.098
-
ATP mutant R199A Bos taurus
2.7.7.19 0.1
-
RNA(A)15 mutant D167N/N202A Bos taurus
2.7.7.19 0.18
-
RNA(A)15 mutant V247R Bos taurus
2.7.7.19 0.26
-
RNA(A)15 mutant D167N/T317G Bos taurus
2.7.7.19 0.308
-
ATP mutant F153A Bos taurus
2.7.7.19 0.39
-
RNA(A)15 mutant V156A Bos taurus
2.7.7.19 0.67
-
RNA(A)15 mutant F100D Bos taurus
2.7.7.19 0.82
-
ATP mutant K158A Bos taurus
2.7.7.19 0.82
-
RNA(A)15 mutant R199A Bos taurus
2.7.7.19 1.055
-
ATP mutant N239A Bos taurus
2.7.7.19 1.073
-
ATP mutant Y237A Bos taurus
2.7.7.19 1.15
-
ATP mutant K228A Bos taurus
2.7.7.19 1.24
-
ATP mutant V154A Bos taurus
2.7.7.19 1.25
-
ATP mutant T317G Bos taurus
2.7.7.19 1.411
-
ATP mutant V247A Bos taurus
2.7.7.19 1.43
-
RNA(A)15 mutant D167N Bos taurus
2.7.7.19 1.476
-
ATP mutant K232A Bos taurus
2.7.7.19 3.19
-
ATP mutant V206A Bos taurus
2.7.7.19 4.203
-
ATP wild-type, reactions are started with the addition of PAP and incubated at 37 °C for 15 minutes. Reactions are stopped by the addition of 8 microL of stop buffer (formamide with trace amount of bromophenol blue dye) Bos taurus
2.7.7.19 4.88
-
ATP mutant Q323A Bos taurus
2.7.7.19 6.092
-
ATP mutant A319R Bos taurus
2.7.7.19 6.76
-
RNA(A)15 mutant K228A Bos taurus
2.7.7.19 6.86
-
RNA(A)15 mutant T317G Bos taurus
2.7.7.19 8.18
-
RNA(A)15 mutant K158A Bos taurus
2.7.7.19 10.52
-
RNA(A)15 mutant F153A Bos taurus
2.7.7.19 11.37
-
RNA(A)15 mutant N239A Bos taurus
2.7.7.19 12.58
-
RNA(A)15 mutant V206A Bos taurus
2.7.7.19 19.25
-
RNA(A)15 mutant Y237A Bos taurus
2.7.7.19 19.71
-
RNA(A)15 mutant Q323A Bos taurus
2.7.7.19 20.34
-
RNA(A)15 mutant K232A Bos taurus
2.7.7.19 22.75
-
RNA(A)15 mutant A319R Bos taurus
2.7.7.19 22.78
-
RNA(A)15 wild-type Bos taurus
2.7.7.19 28.96
-
RNA(A)15 mutant V154A Bos taurus
2.7.7.19 34.5
-
RNA(A)15 mutant V247A Bos taurus