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

BRENDA support

Literature summary for 2.7.2.3 extracted from

  • Hess, D.; Kruger, K.; Knappik, A.; Palm, P.; Hensel, R.
    Dimeric 3-phosphoglycerate kinases from hyperthermophilic Archaea. Cloning, sequencing and expression of the 3-phosphoglycerate kinase gene of Pyrococcus woesei in Escherichia coli and characterization of the protein. Structural and functional comparison with the 3-phosphoglycerate kinase of Methanothermus fervidus (1995), Eur. J. Biochem., 233, 227-237.
    View publication on PubMed
Search for more literature for 2.7.2.3

Cloned(Commentary)

Cloned (Comment) Organism
DNA and amino acid sequence determination and analysis, expression in Escherichia coli JM 83 Pyrococcus woesei
expression in Escherichia coli, the protein expressed in the mesophilic host is folded correctly. By comparing the 3-phosphoglycerate kinase sequences of the mesophilic and the two thermophilic Archaea, trends in thermoadaptation are confirmed that can be deduced from comparisons of glyceraldehyde-3-phosphate dehydrogenase sequences from the same organisms. With increasing temperature the average hydrophobicity and the portion of aromatic residues increases, whereas the chain flexibility as well as the content in chemically labile residues (Asn, Cys) decreases Pyrococcus woesei
expression in Escherichia coli, the protein expressed in the mesophilic host is folded correctly. By comparing the 3-phosphoglycerate kinase sequences of the mesophilic and the two thermophilic Archaea, trends in thermoadaptation are confirmed that can be deduced from comparisons of glyceraldehyde-3-phosphate dehydrogenase sequences from the same organisms. With increasing temperature the average hydrophobicity and the portion of aromatic residues increases, whereas the chain flexibility as well as the content in chemically labile residues (Asn, Cys) decreases Methanothermus fervidus
expression in Escherichia coli. The protein which is expressed in the mesophilic host is folded correctly Pyrococcus woesei
expression in Escherichia coli. The protein which is expressed in the mesophilic host is folded correctly Methanothermus fervidus
expression of slightly modified wild-type in Escherichia coli Methanothermus fervidus

General Stability

General Stability Organism
K+ stabilizes Methanothermus fervidus
K+ stabilizes Pyrococcus woesei
potassium salts of polyvalent anions stabilize the enzyme Pyrococcus woesei
stabilized by anions in decreasing order of efficiency: 2,3-bisphospho-D-glycerate, PO43-, SO42-, Cl- Methanothermus fervidus
the stabilizing effects of anions on thermal stability Pyrococcus woesei
the stabilizing effects of anions on thermal stability Methanothermus fervidus

Inhibitors

Inhibitors Comment Organism Structure
K+ activates Methanothermus fervidus
K+ activates Pyrococcus woesei

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.73
-
 ATP recombinant enzyme, 70°C Methanothermus fervidus
0.73
-
 ATP pH 7.5, 70°C, enzyme expressed in Escherichia coli Methanothermus fervidus
0.73
-
 ATP pH 7.5, 70°C, native enzyme Methanothermus fervidus
0.75
-
 3-phospho-D-glycerate native enzyme, 70°C Methanothermus fervidus
0.75
-
 3-phospho-D-glycerate pH 7.5, 70°C, recombinant enzyme Methanothermus fervidus
0.75
-
 3-phospho-D-glycerate pH 7.5, 70°C, native enzyme Methanothermus fervidus
0.77
-
 3-phospho-D-glycerate recombinant enzyme, 70°C Methanothermus fervidus
0.77
-
 3-phospho-D-glycerate pH 7.5, 70°C, recombinant enzyme Methanothermus fervidus
0.77
-
 3-phospho-D-glycerate pH 7.5, 70°C, enzyme expressed in Escherichia coli Methanothermus fervidus
1.1
-
 Mg2+ native and recombinant enzyme, 70°C Methanothermus fervidus
1.1
-
 ATP native enzyme, 70°C Methanothermus fervidus
1.1
-
 3-phospho-D-glycerate native enzyme, 70°C Pyrococcus woesei
1.1
-
 ATP pH 7.5, 70°C, native enzyme Methanothermus fervidus
1.11
-
 3-phospho-D-glycerate pH 7.5, 70°C, recombinant enzyme Pyrococcus woesei
1.11
-
 ATP pH 7.5, 70°C, enzyme expressed in Escherichia coli Pyrococcus woesei
1.4
-
 3-phospho-D-glycerate recombinant enzyme, 70°C Pyrococcus woesei
1.4
-
 3-phospho-D-glycerate pH 7.5, 70°C, recombinant enzyme Pyrococcus woesei
1.4
-
 3-phospho-D-glycerate pH 7.5, 70°C, enzyme expressed in Escherichia coli Pyrococcus woesei
4.05
-
 ATP native enzyme, 70°C Pyrococcus woesei
4.05
-
 ATP pH 7.5, 70°C, native enzyme Pyrococcus woesei
4.8
-
 ATP recombinant enzyme, 70°C Pyrococcus woesei
4.8
-
 ATP pH 7.5, 70°C, native enzyme Pyrococcus woesei
4.8
-
 3-phospho-D-glycerate pH 7.5, 70°C, native enzyme Pyrococcus woesei

Metals/Ions

Metals/Ions Comment Organism Structure
K+ monovalent cations are essential for the activity of the archaeal enzyme with K+ being significantly more efficient than Na+ Methanothermus fervidus
K+ monovalent cations are essential for the activity of the archaeal enzyme with K+ being significantly more efficient than Na+. Non-cooperative K+ binding with an apparent Kd (K+) of 88 mM Pyrococcus woesei
K+ monovalent cations are essential for the activity, with K+ being significantly more efficient than Na+. Methanothermus fervidus
K+ monovalent cations are essential for the activity, with K+ being significantly more efficient than Na+. Non-cooperative K+ binding with an apparent Kd of 88 mM Pyrococcus woesei
Mg2+
-
 Methanothermus fervidus
Mg2+
-
 Pyrococcus woesei
Mg2+ Km: 1.1 mM Methanothermus fervidus
Na+ monovalent cations are essential for the activity of the archaeal enzyme with K+ being significantly more efficient than Na+ Pyrococcus woesei
Na+ monovalent cations are essential for the activity of the archaeal enzyme with K+ being significantly more efficient than Na+ Methanothermus fervidus
Na+ monovalent cations are essential for the activity, with K+ being significantly more efficient than Na+ Pyrococcus woesei
Na+ monovalent cations are essential for the activity, with K+ being significantly more efficient than Na+ Methanothermus fervidus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
39000
-
 gel filtration, in presence of 0.37 M KCl Methanobacterium bryantii
44800
-
 1 * 44800, SDS-PAGE Methanobacterium bryantii
45000
-
 2 * 45000, SDS-PAGE Methanothermus fervidus
45000
-
 2 * 45000, SDS-PAGE Pyrococcus woesei
45870
-
 2 * 45870, DNA and amino acid sequence determination Methanothermus fervidus
45871
-
 2 * 45871, calculated from sequence Methanothermus fervidus
46161
-
 2 * 46161, DNA and amino acid sequence determination Pyrococcus woesei
46195
-
 2 * 46195, calculated from sequence Pyrococcus woesei
46224
-
 2 * 46224, calculated from sequence Pyrococcus woesei
87000
-
 gel filtration, at 600 mM K+, recombinant enzyme Pyrococcus woesei
87100 90000 recombinant enzyme, gel filtration in presence of 0.6 M KCl, and analytical ultracentrifugation Pyrococcus woesei
90000
-
 ultracentrifugation, recombinant enzyme Pyrococcus woesei
94700
-
 recombinant enzyme, gel filtration in presence of 0.6 M KCl Methanothermus fervidus
94700
-
 gel filtration, at 600 mM K+, recombinant enzyme Methanothermus fervidus
97000
-
 gel filtration, at 600 mM K+, native enzyme Methanothermus fervidus
97100
-
 native enzyme, gel filtration in presence of 0.6 M KCl Methanothermus fervidus
97100
-
 gel filtration at 600 mM K+ Methanothermus fervidus
102700
-
 native enzyme, gel filtration in presence of 0.6 M KCl Pyrococcus woesei
102700
-
 gel filtration at 600 mM K+ Pyrococcus woesei
102700
-
 gel filtration, at 600 mM K+, native enzyme Pyrococcus woesei

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ADP + 3-phospho-D-glyceroyl phosphate Methanothermus fervidus
-
 ATP + 3-phospho-D-glycerate
-
 r
ADP + 3-phospho-D-glyceroyl phosphate Pyrococcus woesei
-
 ATP + 3-phospho-D-glycerate
-
 r
ADP + 3-phospho-D-glyceroyl phosphate Pyrococcus woesei Vul 4 / DSM 3773
-
 ATP + 3-phospho-D-glycerate
-
 r
ADP + 3-phospho-D-glyceroyl phosphate Pyrococcus woesei DSM 3773
-
 ATP + 3-phospho-D-glycerate
-
 r
ATP + 3-phospho-D-glycerate Pyrococcus woesei
-
 ADP + 3-phospho-D-glyceroyl phosphate
-
 ?
ATP + 3-phospho-D-glycerate Methanothermus fervidus
-
 ADP + 3-phospho-D-glyceroyl phosphate
-
 ?
ATP + 3-phospho-D-glycerate Methanothermus fervidus DSM 2088
-
 ADP + 3-phospho-D-glyceroyl phosphate
-
 ?
ATP + 3-phospho-D-glycerate Pyrococcus woesei Vul 4 / DSM 3773
-
 ADP + 3-phospho-D-glyceroyl phosphate
-
 ?
ATP + 3-phospho-D-glycerate Pyrococcus woesei DSM 3773
-
 ADP + 3-phospho-D-glyceroyl phosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Methanobacterium bryantii
-
-
-
Methanothermus fervidus
-
 DSM 2088
-
Methanothermus fervidus P20971
-
-
Methanothermus fervidus DSM 2088 P20971
-
-
Pyrococcus woesei P61884
-
-
Pyrococcus woesei DSM 3773 P61884
-
-
Pyrococcus woesei Vul 4 / DSM 3773 P61884
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Pyrococcus woesei
-
 Methanothermus fervidus
partial Pyrococcus woesei
partial Methanothermus fervidus
partial purification of the native enzyme Methanothermus fervidus
partial purification of the native enzyme Pyrococcus woesei
recombinant wild-type from Escherichia coli JM 83, 97fold Pyrococcus woesei
recombinant, wild-type from Escherichia coli, 51.7fold Methanothermus fervidus

Reaction

Reaction Comment Organism Reaction ID
ATP + 3-phospho-D-glycerate = ADP + 3-phospho-D-glyceroyl phosphate residues engaged in 3-phospho-D-glycerate binding: R22, D24, R144, E149, E150, H191, R192, residues engaged in ATP binding: G236, G237, G263, G371, G375, P372, E277, E377, G407, G408, G429-431 Pyrococcus woesei
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
75
-
 purified recombinant enzyme Methanothermus fervidus
75
-
 pH 7.5, 70°C Pyrococcus woesei
150
-
 purified recombinant enzyme Methanothermus fervidus
150
-
 pH 7.5, 70°C Methanothermus fervidus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ADP + 3-phospho-D-glyceroyl phosphate
-
 Methanothermus fervidus ATP + 3-phospho-D-glycerate
-
 r
ADP + 3-phospho-D-glyceroyl phosphate
-
 Pyrococcus woesei ATP + 3-phospho-D-glycerate
-
 r
ADP + 3-phospho-D-glyceroyl phosphate
-
 Pyrococcus woesei Vul 4 / DSM 3773 ATP + 3-phospho-D-glycerate
-
 r
ADP + 3-phospho-D-glyceroyl phosphate
-
 Pyrococcus woesei DSM 3773 ATP + 3-phospho-D-glycerate
-
 r
ATP + 3-phospho-D-glycerate
-
 Methanobacterium bryantii ADP + 1,3-diphosphoglycerate
-
 r
ATP + 3-phospho-D-glycerate
-
 Methanothermus fervidus ADP + 1,3-diphosphoglycerate
-
 r
ATP + 3-phospho-D-glycerate
-
 Pyrococcus woesei ADP + 1,3-diphosphoglycerate
-
 r
ATP + 3-phospho-D-glycerate
-
 Pyrococcus woesei Vul 4 / DSM 3773 ADP + 1,3-diphosphoglycerate
-
 r
ATP + 3-phospho-D-glycerate
-
 Pyrococcus woesei DSM 3773 ADP + 1,3-diphosphoglycerate
-
 r
ATP + 3-phospho-D-glycerate
-
 Pyrococcus woesei ADP + 3-phospho-D-glyceroyl phosphate
-
 ?
ATP + 3-phospho-D-glycerate
-
 Methanothermus fervidus ADP + 3-phospho-D-glyceroyl phosphate
-
 ?
ATP + 3-phospho-D-glycerate
-
 Methanothermus fervidus DSM 2088 ADP + 3-phospho-D-glyceroyl phosphate
-
 ?
ATP + 3-phospho-D-glycerate
-
 Pyrococcus woesei Vul 4 / DSM 3773 ADP + 3-phospho-D-glyceroyl phosphate
-
 ?
ATP + 3-phospho-D-glycerate
-
 Pyrococcus woesei DSM 3773 ADP + 3-phospho-D-glyceroyl phosphate
-
 ?
ATP + 3-phospho-D-glycerate
-
 Pyrococcus woesei ADP + 3-phospho-D-glyceroyl phosphate.
-
 ?
ATP + 3-phospho-D-glycerate
-
 Pyrococcus woesei Vul 4 / DSM 3773 ADP + 3-phospho-D-glyceroyl phosphate.
-
 ?
ATP + 3-phospho-D-glycerate
-
 Pyrococcus woesei DSM 3773 ADP + 3-phospho-D-glyceroyl phosphate.
-
 ?

Subunits

Subunits Comment Organism
dimer 2 * 45000, SDS-PAGE Methanothermus fervidus
dimer 2 * 45000, SDS-PAGE Pyrococcus woesei
dimer 2 * 45870, DNA and amino acid sequence determination Methanothermus fervidus
dimer 2 * 46161, DNA and amino acid sequence determination Pyrococcus woesei
homodimer 2 * 45000, SDS-PAGE Pyrococcus woesei
homodimer 2 * 45000, SDS-PAGE Methanothermus fervidus
homodimer 2 * 45871, calculated from sequence Methanothermus fervidus
homodimer 2 * 46195, calculated from sequence Pyrococcus woesei
homodimer 2 * 46224, calculated from sequence Pyrococcus woesei
monomer 1 * 44800, SDS-PAGE Methanobacterium bryantii

Synonyms

Synonyms Comment Organism
3-phosphoglycerate kinase
-
 Pyrococcus woesei
3-phosphoglycerate kinase
-
 Methanothermus fervidus
Mfer_0156 ordered locus name Methanothermus fervidus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
70
-
 assay at Methanothermus fervidus
70
-
 assay at Pyrococcus woesei
90
-
-
 Methanothermus fervidus
100
-
 optimum beyond 100°C Pyrococcus woesei
100
-
 temperature optimum beyond 100°C, where activity can not be assayed under the experimental conditions Pyrococcus woesei

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
40 90 the enzyme shows considerable activity at 40°C, 90°C: optimum Methanothermus fervidus
40
-
 only active above 40°C Pyrococcus woesei

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
 stabilizing effects of anions on thermal stability Pyrococcus woesei
additional information
-
 stabilizing effects of anions on thermal stability Methanothermus fervidus
83
-
 half-life: 44 min Methanothermus fervidus
86
-
 half-life: 44 min Methanothermus fervidus
89
-
 half-life: 10 min Methanothermus fervidus
100
-
 half-life: 28 min Pyrococcus woesei
104
-
 half-life: 10 min Pyrococcus woesei

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
113
-
 3-phospho-D-glycerate recombinant enzyme, 70°C Pyrococcus woesei
113.4
-
 ATP pH 7.5, 70°C Pyrococcus woesei
113.4
-
 3-phospho-D-glycerate pH 7.5, 70°C Pyrococcus woesei
113.4
-
 3-phospho-D-glycerate pH 7.5, 70°C, recombinant enzyme Pyrococcus woesei
113.4
-
 ATP pH 7.5, 70°C, native enzyme Pyrococcus woesei
149.8
-
 ATP pH 7.5, 70°C Methanothermus fervidus
149.8
-
 3-phospho-D-glycerate pH 7.5, 70°C Methanothermus fervidus
149.8
-
 ATP pH 7.5, 60°C, native enzyme Methanothermus fervidus
149.8
-
 3-phospho-D-glycerate pH 7.5, 60°C, recombinant enzyme Methanothermus fervidus
150
-
 3-phospho-D-glycerate recombinant enzyme, 70°C Methanothermus fervidus
225.2
-
 ATP pH 7.5, 80°C Methanothermus fervidus
225.2
-
 3-phospho-D-glycerate pH 7.5, 80°C Methanothermus fervidus
225.2
-
 3-phospho-D-glycerate pH 7.5, 70°C, recombinant enzyme Methanothermus fervidus
225.2
-
 ATP pH 7.5, 70°C, native enzyme Methanothermus fervidus
238.5
-
 ATP pH 7.5, 80°C Pyrococcus woesei
238.5
-
 3-phospho-D-glycerate pH 7.5, 80°C Pyrococcus woesei
238.5
-
 3-phospho-D-glycerate pH 7.5, 80°C, recombinant enzyme Pyrococcus woesei
238.5
-
 ATP pH 7.5, 80°C, native enzyme Pyrococcus woesei
254.5
-
 ATP pH 7.5, 90°C Methanothermus fervidus
254.5
-
 3-phospho-D-glycerate pH 7.5, 90°C Methanothermus fervidus
254.5
-
 ATP pH 7.5, 73°C, native enzyme Methanothermus fervidus
254.5
-
 3-phospho-D-glycerate pH 7.5, 73°C, recombinant enzyme Methanothermus fervidus
338.9
-
 3-phospho-D-glycerate pH 7.5, 80°C, recombinant enzyme Methanothermus fervidus
338.9
-
 ATP pH 7.5, 98°C Methanothermus fervidus
338.9
-
 3-phospho-D-glycerate pH 7.5, 98°C Methanothermus fervidus
338.9
-
 ATP pH 7.5, 98°C, native enzyme Methanothermus fervidus
339
-
 3-phospho-D-glycerate recombinant enzyme, 98°C Methanothermus fervidus
505.8
-
 ATP pH 7.5, 90°C Pyrococcus woesei
505.8
-
 3-phospho-D-glycerate pH 7.5, 90°C Pyrococcus woesei
505.8
-
 ATP pH 7.5, 90°C, native enzyme Pyrococcus woesei
505.8
-
 3-phospho-D-glycerate pH 7.5, 90°C, recombinant enzyme Pyrococcus woesei
916
-
 3-phospho-D-glycerate recombinant enzyme, 98°C Pyrococcus woesei
916
-
 ATP pH 7.5, 98°C Pyrococcus woesei
916
-
 3-phospho-D-glycerate pH 7.5, 98°C Pyrococcus woesei
916
-
 ATP pH 7.5, 98°C, native enzyme Pyrococcus woesei
916
-
 3-phospho-D-glycerate pH 7.5, 98°C, recombinant enzyme Pyrococcus woesei

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Pyrococcus woesei
7.5
-
 assay at Methanothermus fervidus

Cofactor

Cofactor Comment Organism Structure
ADP
-
 Methanobacterium bryantii
ADP
-
 Methanothermus fervidus
ADP
-
 Pyrococcus woesei
ATP required as phosphate donor Methanobacterium bryantii
ATP required as phosphate donor Methanothermus fervidus
ATP required as phosphate donor Pyrococcus woesei

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
90
-
 ATP pH 7.5, 93°C Pyrococcus woesei
100
-
 ATP pH 7.5, 73°C Methanothermus fervidus