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Literature summary extracted from

  • Balbo, P.B.; Bohm, A.
    Mechanism of poly(A) polymerase: Structure of the enzyme-MgATP-RNA ternary complex and kinetic analysis (2007), Structure, 15, 1117-1131.
    View publication on PubMedView publication on EuropePMC

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.7.19 mutant D154A in complex with MgATP-RNA, hanging drop vapour diffusion method, in 0.1 M bis-Tris propane, pH 6.4, 0.2 M Li-acetate, and 16% PEG 3350 Saccharomyces cerevisiae
2.7.7.19 mutant D154A, trapped in complex with ATP and a five residue poly(A). Enzyme has undergone significant domain movement and shows a closed conformation with extensive interactions between substrates and all three polymerase domains Saccharomyces cerevisiae

Protein Variants

EC Number Protein Variants Comment Organism
2.7.7.19 D154A crystallization data, closed conformation with extensive interactions between substrates and all three polymerase domains Saccharomyces cerevisiae
2.7.7.19 D154A the mutant has a nearly identical melting temperature as wild type PAP Saccharomyces cerevisiae
2.7.7.19 K215A 2- to 4fold increase in Km values Saccharomyces cerevisiae
2.7.7.19 K215A the mutant has a nearly identical melting temperature as wild type PAP Saccharomyces cerevisiae
2.7.7.19 N189A residue bridges the N and middle domains in the closed state Saccharomyces cerevisiae
2.7.7.19 N189A the mutant has a nearly identical melting temperature as wild type PAP Saccharomyces cerevisiae
2.7.7.19 N226A mutation affects the equilibrium between the open- and closed-domain forms of the enzyme Saccharomyces cerevisiae
2.7.7.19 N226A the mutant has a nearly identical melting temperature as wild type PAP Saccharomyces cerevisiae
2.7.7.19 Y224F mutation affects the equilibrium between the open- and closed-domain forms of the enzyme Saccharomyces cerevisiae
2.7.7.19 Y224F altered melting temperature (44°C) compared to the wild type enzyme Saccharomyces cerevisiae

General Stability

EC Number General Stability Organism
2.7.7.19 substrates poly(A) and Mg-ATP induce the conformational change, resulting in stabilization of the closed enzyme state and enabling catalysis Saccharomyces cerevisiae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.7.19 0.036
-
 ATP wild-type, pH 7.0, 30°C Saccharomyces cerevisiae
2.7.7.19 0.0468
-
 (A)n wild-type, pH 7.0, 30°C Saccharomyces cerevisiae
2.7.7.19 0.055
-
 GTP mutant N226A, pH 7.0, 30°C Saccharomyces cerevisiae
2.7.7.19 0.062
-
 GTP wild-type, pH 7.0, 30°C Saccharomyces cerevisiae
2.7.7.19 0.08
-
 ATP mutant N189A, pH 7.0, 30°C Saccharomyces cerevisiae
2.7.7.19 0.104
-
 CTP wild-type, pH 7.0, 30°C Saccharomyces cerevisiae
2.7.7.19 0.106
-
 (A)n mutant N189A, pH 7.0, 30°C Saccharomyces cerevisiae
2.7.7.19 0.14
-
 CTP mutant N226A, pH 7.0, 30°C Saccharomyces cerevisiae
2.7.7.19 0.148
-
 CTP mutant N189A, pH 7.0, 30°C Saccharomyces cerevisiae
2.7.7.19 0.195
-
 (A)n mutant K215A, pH 7.0, 30°C Saccharomyces cerevisiae
2.7.7.19 0.249
-
 ATP mutant N226A, pH 7.0, 30°C Saccharomyces cerevisiae
2.7.7.19 0.367
-
 (A)n mutant N226A, pH 7.0, 30°C Saccharomyces cerevisiae
2.7.7.19 0.368
-
 CTP mutant K215A, pH 7.0, 30°C Saccharomyces cerevisiae
2.7.7.19 0.406
-
 ATP mutant K215A, pH 7.0, 30°C Saccharomyces cerevisiae
2.7.7.19 0.711
-
 (A)n mutant Y224F, pH 7.0, 30°C Saccharomyces cerevisiae
2.7.7.19 0.929
-
 ATP mutant Y224F, pH 7.0, 30°C Saccharomyces cerevisiae
2.7.7.19 4.7
-
 CTP mutant Y224F, pH 7.0, 30°C Saccharomyces cerevisiae

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.7.19 Mg2+ essential for activity Saccharomyces cerevisiae

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.19 Saccharomyces cerevisiae P29468
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.7.19 nickel affinity column chromatography and ion exchange chromatography Saccharomyces cerevisiae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.19 (A)n + ATP
-
 Saccharomyces cerevisiae (A)n+1 + diphosphate
-
 ?
2.7.7.19 (A)n + CTP
-
 Saccharomyces cerevisiae (A)n-C + diphosphate
-
 ?
2.7.7.19 (A)n + diphosphate
-
 Saccharomyces cerevisiae (A)n-1 + ATP
-
 ?
2.7.7.19 (A)n + GTP
-
 Saccharomyces cerevisiae (A)n-G + diphosphate
-
 ?
2.7.7.19 ATP + RNA
-
 Saccharomyces cerevisiae diphosphate + RNA(A)n
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.7.7.19 PAP PAP is a template-independent polymerase that belongs to the DNA polymerase beta family of enzymes Saccharomyces cerevisiae
2.7.7.19 poly(A) polymerase
-
 Saccharomyces cerevisiae

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.7.7.19 30 48 remains stable at 30°C, the melting temperature for the wild type enzyme is at 48°C Saccharomyces cerevisiae