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Literature summary for 2.7.2.1 extracted from

  • Gorrell, A.; Ferry, J.G.
    Investigation of the Methanosarcina thermophila acetate kinase mechanism by fluorescence quenching (2007), Biochemistry, 46, 14170-14176.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.7.2.1

Cloned(Commentary)

Cloned (Comment) Organism
overexpressed in Escherichia coli BL21(DE3) Methanosarcina thermophila

Protein Variants

Protein Variants Comment Organism
Q43W site-directed mutagenesis, single mutant Methanosarcina thermophila
R241A/Q43W site-directed mutagenesis, double mutant Methanosarcina thermophila
R241K/Q43W site-directed mutagenesis, double mutant Methanosarcina thermophila
R91A/Q43W site-directed mutagenesis, double mutant Methanosarcina thermophila
R91K/Q43W site-directed mutagenesis, double mutant Methanosarcina thermophila

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.053
-
 ATP Q43W, mutant Methanosarcina thermophila
0.08
-
 ATP wild-type Methanosarcina thermophila
0.08
-
 ATP R241A/Q43W, double mutant Methanosarcina thermophila
0.098
-
 ADP wild-type Methanosarcina thermophila
0.147
-
 ADP R91A/Q43W, double mutant Methanosarcina thermophila
0.164
-
 ADP R91K/Q43W, double mutant Methanosarcina thermophila
0.173
-
 ADP Q43W, mutant Methanosarcina thermophila
0.234
-
 ATP R91K/Q43W, double mutant Methanosarcina thermophila
0.239
-
 ADP R241A/Q43W, double mutant Methanosarcina thermophila
0.26
-
 acetyl phosphate Q43W Methanosarcina thermophila
0.33
-
 acetyl phosphate R241/Q43W Methanosarcina thermophila
0.47
-
 acetyl phosphate wild-type Methanosarcina thermophila
0.539
-
 ATP R91A/Q43W, double mutant Methanosarcina thermophila
0.692
-
 ADP R241K/Q43W Methanosarcina thermophila
1.02
-
 acetyl phosphate R241K/Q43W Methanosarcina thermophila
1.08
-
 acetyl phosphate R91K/Q43W Methanosarcina thermophila
1.3
-
 ATP R241K/Q43W, double mutant Methanosarcina thermophila
2.58
-
 acetyl phosphate R91A/Q43W Methanosarcina thermophila
2.7
-
 acetate wild-type Methanosarcina thermophila
5.8
-
 acetate R241A/Q43W, double mutant Methanosarcina thermophila
12.4
-
 acetate Q43W, mutant Methanosarcina thermophila
48
-
 acetate R91K/Q43W, double mutant Methanosarcina thermophila
179
-
 acetate R241K/Q43W, double mutant Methanosarcina thermophila

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ binding constants of enzyme variants with ADP measured Methanosarcina thermophila

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + acetate Methanosarcina thermophila
-
 ADP + acetyl phosphate
-
 r

Organism

Organism UniProt Comment Textmining
Methanosarcina thermophila P38502
-
-

Purification (Commentary)

Purification (Comment) Organism
judged by SDS-PAGE Methanosarcina thermophila

Reaction

Reaction Comment Organism Reaction ID
ATP + acetate = ADP + acetyl phosphate roles for the arginine residues Arg241 and Arg91 in transition state stabilization for catalysis but not in nucleotide binding determined, experimental evidence for domain motion Methanosarcina thermophila
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 catalytic mechanism analyzed in wild-type and mutant variants, binding constants for the nucleotide substrates indicate that Arg241 is involved in transition state stabilization and not directly involved in nucleotide recognition or binding, or in the domain closure required for catalysis, binding constants of the nucleotide substrates for Arg91 suggest that this residue has a role in transition state stabilization, evidence for domain motion dependent upon nucleotide ligand binding presented, suggestion that Arg91 is important for closure of domain I onto domain II for catalysis Methanosarcina thermophila

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + acetate
-
 Methanosarcina thermophila ADP + acetyl phosphate
-
 r
ATP + acetate catalytic mechanism, roles for the arginine residues Arg241 and Arg91 in transition state stabilization for catalysis but not in nucleotide binding determined, experimental evidence for domain motion Methanosarcina thermophila ADP + acetyl phosphate
-
 r

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.4
-
 ATP R241K/Q43W, double mutant Methanosarcina thermophila
1.83
-
 ATP R241A/Q43W, double mutant Methanosarcina thermophila
2.07
-
 ATP R91A/Q43W, double mutant Methanosarcina thermophila
2.63
-
 ADP R91A/Q43W, double mutant Methanosarcina thermophila
10.5
-
 ADP R241K/Q43W, double mutant Methanosarcina thermophila
11.8
-
 ATP R91K/Q43W, double mutant Methanosarcina thermophila
14.6
-
 ADP R241A/Q43W, double mutant Methanosarcina thermophila
25.7
-
 ADP R91K/Q43W, double mutant Methanosarcina thermophila
699
-
 ATP Q43W, mutant Methanosarcina thermophila
913
-
 ATP wild-type Methanosarcina thermophila
1255
-
 ADP wild-type Methanosarcina thermophila
2650
-
 ADP Q43W, mutant Methanosarcina thermophila

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Methanosarcina thermophila