Protein Variants | Comment | Organism |
---|---|---|
DELTA1-398 | the autoinhibitory domain is separately expressed and purified. Trans-addition of purified AID effectively inhibits PDK1-catalyzed T-loop phosphorylation of a catalytic kinase domain construct of S6K1 | Homo sapiens |
DELTA1-398/S411D/S418D/T421E/S424D | expression of the autoinhibitory domain with several mutations shows that the tetraphospho-mimicking mutations in the autoinhibitory domain construct significantly decreased AID-mediated inhibition of PDK1-catalyzed T229 phosphorylation of DELTA399-502/T389E (His6-S6K1(DELTA AID)-T389E) | Homo sapiens |
DELTA399-502/T389E | by using a mutant protein wich lacks the autoinhibitory domain (AID) and which contains a T389E mutation it is shown that the autoinhibiotry domain (AID) of S6K1 expressed separately most significantly inhibits PDK1-catalyzed T229 phosphorylation in the T-loop of the S6K1 catalytic kinase domain construct, His6-S6K1(DELTA AID)-T389E | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
auoinhibitory domain of S6K1 | - |
Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P23443 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
phosphoprotein | catalytic activation requires dual phosphorylation of critical residues in the conserved T-loop (T229) and hydrophobic motif (HM, T389) peptide regions of its catalytic kinase domain (residues 1-398) | Homo sapiens |
Purification (Comment) | Organism |
---|---|
optimal bacterial expression and affinity purification of the full autoinhibitory domain (AID) of S6K1 is achieved by fusing GST to the N-terminus, growing the BL21 (DE3) strain of Escherichia coli in PG minimal medium, adding the irreversible protease inhibitors TPCK and TLCK to the lysis buffer, using the highly selective PreScission Protease to remove the GST affinity tag, and using the Superdex 75 size-exclusion column to resolve cleaved full length AID from smaller proteolyzed fragments | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
S6K1 | - |
Homo sapiens |
serine-threonine protein kinase | - |
Homo sapiens |