Competitive ligands facilitate dissociation of the complex of bifunctional inhibitor and protein kinase

Ivan, Taavi; Enkvist, Erki; Sinijärv, Hedi; Uri, Asko

doi:10.1016/j.bpc.2017.06.004

Cited by 5 publications

(6 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The dissociation rate of the reversible inhibitors (6−9) from their complex with PKAcα was measured by the following protocol of the TGLI assay. 42 At first, solution of the inhibitor and recombinant PKAcα protein was pre-incubated for 1 h to form the complex. Thereafter, the photoluminescent probe was added in great excess to completely displace the inhibitor and the time course of the displacement was monitored by TGLI measurements (Figure 3b, Table 1).…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Construction of Covalent Bisubstrate Inhibitor of Protein Kinase Reacting with Cysteine Residue at Substrate-Binding Site

et al. 2022

Self Cite

View full text Add to dashboard Cite

Recent clinical success with targeted covalent inhibitors points to new possibilities for development of protein kinase (PK)-targeted drugs by exploiting reactive cysteine residues in and around the ATP-binding site. However, more than 300 human PKs lack cysteine residues in the ATP-binding site. Here, we report the first covalent bisubstrate PK inhibitor whose electrophilic warhead reaches outside the ATP-binding site and reacts with a distant cysteine residue. A series of covalent inhibitors and their reversible counterparts were synthesized and characterized. The most potent reversible inhibitor possessed picomolar affinity and its cysteine-reactive counterpart revealed high value of k inact/K I ratio (6.2 × 107 M–1 s–1) for the reaction with the catalytic subunit of cAMP-dependent PK (PKAc). Under optimized conditions, fluorescent dye-labeled covalent inhibitors demonstrated PKA-selectivity in the cell lysate and reacted with several proteins inside live cells, including PKAc. The disclosed compounds serve as leads for targeting PKs possessing an analogously positioned cysteine residue.

show abstract

Section: ■ Results and Discussionmentioning

confidence: 99%

Construction of Covalent Bisubstrate Inhibitor of Protein Kinase Reacting with Cysteine Residue at Substrate-Binding Site

et al. 2022

Self Cite

View full text Add to dashboard Cite

show abstract

“…As a result, the dissociated moiety cannot rebind to the receptor and the multivalent ligand completely dissociates from the ligand-receptor complexes. In other words, if the dissociation kinetics is accelerated in the presence of a competitor, one can assume that the multivalent ligand forms multiple bonds. − …”

Section: Discussionmentioning

confidence: 99%

Influence of Linker Molecules in Hexavalent RGD Peptides on Their Multivalent Interactions with Integrin α_vβ₃

et al. 2021

View full text Add to dashboard Cite

Multivalent RGD peptides have been used as an excellent targeting vector to integrin α v β 3 -positive tumors. However, little attention has been paid to the influence of linker molecules in multivalent RGD peptides on their dissociation kinetics from tumor cells. In this study, we evaluated the dissociation kinetics of 99m Tc-labeled hexavalent RGD peptides which have (CH 2 -CH 2 -O) n (n = 4, [ 99m Tc][Tc(L1) 6 ] + and n = 12, [ 99m Tc][Tc(L2) 6 ] + ) or ( D Pro-Gly) n (n = 1, [ 99m Tc][Tc(L3) 6 ] + ; n = 6, [ 99m Tc][Tc(L4) 6 ] + ; and n = 9, [ 99m Tc][Tc(L5) 6 ] + ) as a linker molecule. The results showed that [ 99m Tc][Tc(L4) 6 ] + and [ 99m Tc][Tc(L5) 6 ] + displayed slower dissociation kinetics and [ 99m Tc][Tc(L4) 6 ] + showed exceptionally high in vitro cellular uptake (203.1 ± 16.7% dose/mg protein) and the highest tumor to blood ratio (138.1 ± 26.3 at 4 h p.i.) in tumor bearing nude mice. These findings indicate that the use of appropriate length of ( D Pro-Gly) n would maximize the binding of multivalent RGD peptides to clustered integrin α v β 3 .

show abstract

“…Finally, bisubstrate-analog inhibitors incorporate fragments that can bind to both ATP-and substrate-binding site of PK, and hence need to compete with intracellular ATP as well as endogenous substrate for binding. While it is possible that only one fragment of the bisubstrate-analog inhibitor gets bound to the suitable site in PK molecule [166], simultaneous binding of both fragments is thermodynamically favored. Therefore, bisubstrate-analog inhibitors bind preferentially to PKs residing in sufficiently open conformation.…”

Section: Class 3amentioning

confidence: 99%

Dissection of Protein Kinase Pathways in Live Cells Using Photoluminescent Probes: Surveillance or Interrogation?

2018

View full text Add to dashboard Cite

Protein kinases catalyze phosphorylation, a small yet crucial modification that affects participation of the substrate proteins in the intracellular signaling pathways. The activity of 538 protein kinases encoded in human genome relies upon spatiotemporally controlled mechanisms, ensuring correct progression of virtually all physiological processes on the cellular level-from cell division to cell death. The aberrant functioning of protein kinases is linked to a wide spectrum of major health issues including cancer, cardiovascular diseases, neurodegenerative diseases, inflammatory diseases, etc. Hence, significant effort of scientific community has been dedicated to the dissection of protein kinase pathways in their natural milieu. The combination of recent advances in the field of light microscopy, the wide variety of genetically encoded or synthetic photoluminescent scaffolds, and the techniques for intracellular delivery of cargoes has enabled design of a plethora of probes that can report activation of target protein kinases in human live cells. The question remains: how much do we bias intracellular signaling of protein kinases by monitoring it? This review seeks answers to this question by analyzing different classes of probes according to their general structure, mechanism of recognition of biological target, and optical properties necessary for the reporting of intracellular events.

show abstract

Competitive ligands facilitate dissociation of the complex of bifunctional inhibitor and protein kinase

Cited by 5 publications

References 32 publications

Construction of Covalent Bisubstrate Inhibitor of Protein Kinase Reacting with Cysteine Residue at Substrate-Binding Site

Construction of Covalent Bisubstrate Inhibitor of Protein Kinase Reacting with Cysteine Residue at Substrate-Binding Site

Influence of Linker Molecules in Hexavalent RGD Peptides on Their Multivalent Interactions with Integrin α_vβ₃

Dissection of Protein Kinase Pathways in Live Cells Using Photoluminescent Probes: Surveillance or Interrogation?

Contact Info

Product

Resources

About

Competitive ligands facilitate dissociation of the complex of bifunctional inhibitor and protein kinase

Cited by 5 publications

References 32 publications

Construction of Covalent Bisubstrate Inhibitor of Protein Kinase Reacting with Cysteine Residue at Substrate-Binding Site

Construction of Covalent Bisubstrate Inhibitor of Protein Kinase Reacting with Cysteine Residue at Substrate-Binding Site

Influence of Linker Molecules in Hexavalent RGD Peptides on Their Multivalent Interactions with Integrin αvβ3

Dissection of Protein Kinase Pathways in Live Cells Using Photoluminescent Probes: Surveillance or Interrogation?

Contact Info

Product

Resources

About

Influence of Linker Molecules in Hexavalent RGD Peptides on Their Multivalent Interactions with Integrin α_vβ₃