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Literature summary for 2.7.11.1 extracted from

  • Keshwani, M.M.; Ross, D.B.; Ragan, T.J.; Harris, T.K.
    Baculovirus-mediated expression, purification, and characterization of a fully activated catalytic kinase domain construct of the 70 kDa 40S ribosomal protein S6 kinase-1 alphaII isoform (S6K1alphaII) (2008), Protein Expr. Purif., 58, 32-41.
    View publication on PubMedView publication on EuropePMC

Protein Variants

Protein Variants Comment Organism
DELTA399-502/T389E an N-terminal His6 affinity tagged catalytic kinase domain construct of the alphaII isoform of S6K1 lacking the autoinhibitory domain (AID) (His6-S6K1alphaII(DELTA AID)-T389E), is generated in its most highly active form (250 nmol/min/mg) by baculovirus-mediated expression and purification from Sf9 insect cells that are coinfected with recombinant PDK1. kcat (1/sec): 0.19 (RRRLSSLRA) in vivo assay, 0.057 (RRRLSSLRA) in vitro assay, 0.19 (ATP) in vivo assay, 0.057 (ATP) in vitro assay. Km (mM): 3.6 (RRRLSSLRA) in vivo assay, 3.7 (RRRLSSLRA) in vitro assay, 5.4 (ATP) in vivo assay, 4.8 (ATP) in vitro assay Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0036
-
RRRLSSLRA in vivo PDK1-activated mutant DELTA399-502/T389E (His6-S6K1aII(DELTA AID)-T389E) Homo sapiens
0.0037
-
RRRLSSLRA in vitro PDK1-activated mutant DELTA399-502/T389E (His6-S6K1aII(DELTA AID)-T389E) Homo sapiens
0.0048
-
ATP in vitro PDK1-activated mutant DELTA399-502/T389E (His6-S6K1aII(DELTA AID)-T389E) Homo sapiens
0.0054
-
ATP in vivo PDK1-activated mutant DELTA399-502/T389E (His6-S6K1aII(DELTA AID)-T389E) Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
an N-terminal His6 affinity tagged catalytic kinase domain construct of the alphaII isoform of S6K1 lacking the autoinhibitory domain (AID) (His6-S6K1alphaII(DELTA AID)-T389E), is expressed in Escherichia coli and purified by His6 affinity co-purification and by MonoQ anion exchange chromatography at pH 7.3 Homo sapiens

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.25
-
specific activity of an N-terminal His6 affinity tagged catalytic kinase domain construct of the alphaII isoform of S6K1 lacking the autoinhibitory domain (AID) (DELTA399-502/T389E) Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + RRRLSSLRA enzyme activities are determined by measuring initial velocities for His6-S6K1aII(DELTA AID)-T389E catalyzed phosphorylation of a commercially available S6K/RSK model peptide substrate 1 (S6K1-Tide, RRRLSSLRA) Homo sapiens ADP + RRRLSpSLRA
-
?

Synonyms

Synonyms Comment Organism
S6K1alphaII
-
Homo sapiens
serine-threonine kinase
-
Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.057
-
ATP in vitro PDK1-activated mutant DELTA399-502/T389E (His6-S6K1aII(DELTA AID)-T389E) Homo sapiens
0.057
-
RRRLSSLRA in vitro PDK1-activated mutant DELTA399-502/T389E (His6-S6K1aII(DELTA AID)-T389E) Homo sapiens
0.19
-
ATP in vivo PDK1-activated mutant DELTA399-502/T389E (His6-S6K1aII(DELTA AID)-T389E) Homo sapiens
0.19
-
RRRLSSLRA in vivo PDK1-activated mutant DELTA399-502/T389E (His6-S6K1aII(DELTA AID)-T389E) Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Homo sapiens