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Literature summary for 2.7.1.25 extracted from

  • Sekulic, N.; Konrad, M.; Lavie, A.
    Structural mechanism for substrate inhibition of the adenosine 5-phosphosulfate kinase domain of human 3-phosphoadenosine 5-phosphosulfate synthetase 1 and its ramifications for enzyme regulation (2007), J. Biol. Chem., 282, 22112-22121.
    View publication on PubMed
Search for more literature for 2.7.1.25

Cloned(Commentary)

Cloned (Comment) Organism
APS kinase domain (amino acids 25-227) of the PAPSS1 coding region are expressed as a GST-fusion protein in Escherichia coli Homo sapiens

Crystallization (Commentary)

Crystallization (Comment) Organism
-
 Homo sapiens
crystals are grown at room temperature using hanging drops containing equal volumes of protein and reservoir solution Homo sapiens

Protein Variants

Protein Variants Comment Organism
delta34N deletion mutant lacking the first 34 N-terminal residues from APS kinase domain results in an enzyme with similar kinetic behaviour to the full-length domain Homo sapiens
delta50N deletion mutant lacking the first 50 N-terminal residues from APS kinase domain show no substrate inhibition by adenylyl sulfate and approximately half of the full-length's turnover number, crystal structure reveals an asymmetrical dimer Homo sapiens
additional information the results show that the alpha1 Helix constructed by N-terminal residue 35-50 of the APS kinase domain are critical in stabilizing a symmetrical dimer, interactions established by the highly conserved arginines 37 and 40 are indispensable for maintaining substrate inhibition of the APS kinase domain in human PAPSS1 Homo sapiens
additional information generation of truncated and point mutants of the APS kinase domain that are active but devoid of substrate inhibition. Structural analysis of these mutant enzymes reveals the intrasubunit rearrangements that occur upon substrate binding Homo sapiens
R37A mutant clone shows no substrate inhibition by adenylyl sulfate, mutant is kinetically indistinguishable from deletion mutant delta50N lacking the first N-terminal residues Homo sapiens
R40A mutant clone shows no substrate inhibition by adenylyl sulfate, mutant is kinetically indistinguishable from deletion mutant delta50N lacking the first N-terminal residues Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
adenosine 5'-phosphosulfate acts as a strong uncompetitive inhibitor of the APS kinase reaction Homo sapiens
adenylyl sulfate adenylyl sulfate acts as a strong uncompetitive inhibitor of the APS kinase reaction Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.002
-
 adenylyl sulfate value is below 0.002 for a deletion mutant lacking the first 34 amino acids of the N-terminus, delta34N Homo sapiens
0.002
-
 adenylyl sulfate value is below 0.002 for wildtype adenylyl sulfate Homo sapiens
0.0032
-
 adenylyl sulfate mutant R37A Homo sapiens
0.0037
-
 adenylyl sulfate mutant R40A Homo sapiens
0.0046
-
 adenylyl sulfate deletion mutant lacking the first 50 amino acids of the N-terminus, delta50N Homo sapiens

Metals/Ions

Metals/Ions Comment Organism Structure
EDTA assay with 0.25 mM EDTA Homo sapiens
K+ assay with 100 mM KCl Homo sapiens
Mg2+ assay with 2.5 mM MgCl2 Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens O43252
-
-
Homo sapiens O95340
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Homo sapiens
protein solutions are applied on Blue-Sepharose columns and further purified by ion-exchange chromatography Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
MgATP2- + adenylyl sulfate
-
 Homo sapiens MgADP- + 3'-phosphoadenylyl sulfate
-
 r

Synonyms

Synonyms Comment Organism
3'-phosphoadenosine 5'-phosphosulfate synthetase1
-
 Homo sapiens
adenosine 5-phosphosulfate kinase adenosine 5-phosphosulfate kinase is a domain of human 3-phosphoadenosine 5-phosphosulfate synthetase 1 Homo sapiens
PAPSS1
-
 Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.17
-
 adenylyl sulfate mutant R40A Homo sapiens
0.2
-
 adenylyl sulfate deletion mutant lacking the first 50 amino acids of the N-terminus, delta50N Homo sapiens
0.207
-
 adenylyl sulfate mutant R37A Homo sapiens
0.315
-
 adenylyl sulfate deletion mutant lacking the first 34 amino acids of the N-terminus, delta34N Homo sapiens
0.364
-
 adenylyl sulfate wildtype adenylyl sulfate Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Homo sapiens

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
additional information
-
 adenylyl sulfate no inhibition of a deletion mutant lacking the first 50 amino acids of the N-terminus, delta50N Homo sapiens
additional information
-
 adenylyl sulfate no inhibition of mutant R37A Homo sapiens
additional information
-
 adenylyl sulfate no inhibition of mutant R40A Homo sapiens
0.017
-
 adenylyl sulfate wild type adenylyl sulfate Homo sapiens
0.029
-
 adenylyl sulfate deletion mutant lacking the first 34 amino acids of the N-terminus, delta34N Homo sapiens