scholarly journals Structural basis for UCN-01 (7-hydroxystaurosporine) specificity and PDK1 (3-phosphoinositide-dependent protein kinase-1) inhibition

2003 ◽  
Vol 375 (2) ◽  
pp. 255-262 ◽  
Author(s):  
David KOMANDER ◽  
Gursant S. KULAR ◽  
Jennifer BAIN ◽  
Matthew ELLIOTT ◽  
Dario R. ALESSI ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Hydroxy Group ◽  
Active Site ◽  
Binding Pocket ◽  
Kinase Domain ◽  
Structural Basis ◽  
Active Site Residues ◽  
Dependent Protein Kinase ◽  
Growth Factor Signalling ◽  
Dependent Protein

PDK1 (3-phosphoinositide-dependent protein kinase-1) is a member of the AGC (cAMP-dependent, cGMP-dependent, protein kinase C) family of protein kinases, and has a key role in insulin and growth-factor signalling through phosphorylation and subsequent activation of a number of other AGC kinase family members, such as protein kinase B. The staurosporine derivative UCN-01 (7-hydroxystaurosporine) has been reported to be a potent inhibitor for PDK1, and is currently undergoing clinical trials for the treatment of cancer. Here, we report the crystal structures of staurosporine and UCN-01 in complex with the kinase domain of PDK1. We show that, although staurosporine and UCN-01 interact with the PDK1 active site in an overall similar manner, the UCN-01 7-hydroxy group, which is not present in staurosporine, generates direct and water-mediated hydrogen bonds with active-site residues. Inhibition data from UCN-01 tested against a panel of 29 different kinases show a different pattern of inhibition compared with staurosporine. We discuss how these differences in inhibition could be attributed to specific interactions with the additional 7-hydroxy group, as well as the size of the 7-hydroxy-group-binding pocket. This information could lead to opportunities for structure-based optimization of PDK1 inhibitors.

Substrate-mediated channeling of a chemical reagent to the active site of cAMP-dependent protein kinase

FEBS Letters ◽  
1981 ◽  
Vol 130 (1) ◽  
pp. 127-132 ◽  
Author(s):  
Juan S. Jiménez ◽  
Abraham Kupfer ◽  
Philip Gottlieb ◽  
Shmuel Shaltiel
Keyword(s):  
Protein Kinase ◽  
Active Site ◽  
Chemical Reagent ◽  
Dependent Protein Kinase ◽  
Camp Dependent Protein Kinase ◽  
Dependent Protein

scholarly journals Structural basis of cyclic nucleotide selectivity in cGMP dependent protein kinase II

2015 ◽  
Vol 16 (S1) ◽  
Author(s):  
James C. Campbell ◽  
Kevin Y. Li ◽  
Jeong Joo Kim ◽  
Gilbert Huang ◽  
Albert S. Reger ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Cyclic Nucleotide ◽  
Structural Basis ◽  
Dependent Protein Kinase ◽  
Cgmp Dependent Protein Kinase ◽  
Protein Kinase Ii ◽  
Dependent Protein

scholarly journals Insights into the phosphoryl transfer catalyzed by cAMP-dependent protein kinase

2014 ◽  
Vol 70 (a1) ◽  
pp. C449-C449
Author(s):  
Oksana Gerlits ◽  
Amit Das ◽  
Jianhui Tian ◽  
Malik Keshwani ◽  
Susan Taylor ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Protein Kinases ◽  
Active Site ◽  
Phosphate Group ◽  
Nucleoside Triphosphate ◽  
Phosphoryl Transfer ◽  
Hydrogen Atoms ◽  
Dependent Protein Kinase ◽  
Camp Dependent Protein Kinase ◽  
Dependent Protein

Protein kinases are involved in a number of cell signaling pathways. They catalyze phosphorylation of proteins and regulate the majority of cellular processes (such as growth, differentiation, lipid metabolism, regulation of sugar, nucleic acid synthesis, etc.). Chemically, protein kinases covalently transfer the gamma-phosphate group of a nucleoside triphosphate (e.g. ATP) to a hydroxyl group of a Ser, Thr or Tyr residue of substrate protein or peptide. The reaction involves moving hydrogen atoms between the enzyme, substrate and nucleoside. The unanswered question is whether the proton transfer from the Ser residue happens before the phosphoryl transfer using the general acid-base catalyst, Asp166, or after the reaction went through the transition state by directly protonating the phosphate group. To address this key question about the phosphoryl transfer, we determined a number of X-ray structures of ternary complexes of catalytic subunit of cAMP-dependent protein kinase (PKAc) with various substrates, nucleotides and cofactors. Importantly, we were able to trap and mimic the initial (Michaelis complex) and final (product complex) stages of the reaction. The results demonstrate that Mg2+, Ca2+, Sr2+, and Ba2+ metal ions bind to the active site and facilitate the reaction to produce ADP and a phosphorylated peptide. The study also revealed that metal-free PKAc can facilitate the phosphoryl transfer reaction; a result that was confirmed with single turnover enzyme kinetics measurements. Comparison of the product and the pseudo-Michaelis complex structures, in conjunction with molecular dynamics simulations, reveals conformational, coordination, and hydrogen bonding changes that help further our understanding of the mechanism, roles of metals, and active site residues involved in PKAc activity.

scholarly journals Exploiting the Unique ATP-Binding Pocket ofToxoplasmaCalcium-Dependent Protein Kinase 1 To Identify Its Substrates

2013 ◽  
Vol 8 (6) ◽  
pp. 1155-1162 ◽  
Author(s):  
Sebastian Lourido ◽  
Grace R. Jeschke ◽  
Benjamin E. Turk ◽  
L. David Sibley
Keyword(s):  
Protein Kinase ◽  
Binding Pocket ◽  
Atp Binding ◽  
Dependent Protein Kinase ◽  
Dependent Protein
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