A synthetic peptide that prevents camp regulation in mammalian hyperpolarization-activated cyclic nucleotide-gated (HCN) channels
Articolo
Data di Pubblicazione:
2018
Abstract:
Binding of TRIP8b to the cyclic nucleotide binding domain (CNBD) of mammalian hyperpolarization-activated cyclic nucleotide-gated (HCN) channels prevents their regulation by cAMP. Since TRIP8b is expressed exclusively in the brain, we envisage that it can be used for orthogonal control of HCN channels beyond the central nervous system. To this end, we have identified by rational design a 40-aa long peptide (TRIP8b nano ) that recapitulates affinity and gating effects of TRIP8b in HCN isoforms (hHCN1, mHCN2, rbHCN4) and in the cardiac current I f in rabbit and mouse sinoatrial node cardiomyocytes. Guided by an NMR-derived structural model that identifies the key molecular interactions between TRIP8b nano and the HCN CNBD, we further designed a cell-penetrating peptide (TAT-TRIP8b nano ) which successfully prevented ?-adrenergic activation of mouse I f leaving the stimulation of the L-type calcium current (I CaL ) unaffected. TRIP8b nano represents a novel approach to selectively control HCN activation, which yields the promise of a more targeted pharmacology compared to pore blockers. © Saponaro et al.
Tipologia CRIS:
01.01 Articolo in rivista
Keywords:
asparagine; cell penetrating peptide; cyclic AMP; hyperpolarization activated cyclic nucleotide gated channel; isoprenaline; synthetic peptide; calcium channel L type; cyclic AMP; HCN1 protein; human; Hcn2 protein; mouse; hyperpolariza; membrane protein; peptide; peroxin; Pex5l protein; mouse; potassium channel; protein binding; transactivator protein; action potential; animal cell; Article; binding affinity; calcium current; carboxy terminal sequence; cardiac muscle cell; channel gating; controlled study; current density; deletion mutant; female; half-activation voltage; heart automaticity; heteronuclear single quantum coherence; human; hydrogen bond; isothermal titration calorimetry; male; molecular docking; molecular interaction; mouse; nonhuman; nuclear magnetic resonance spectroscopy; protein secondary structure; sinus node; alpha helix; animal; beta sheet; binding site; C57BL mouse; chemistry; cytology; drug effect; gene expression; genetics; HEK293 cell line; Leporidae; metabolism; patch clamp technique; protein domain; synthesis; Animals; Binding Sites; Calcium Channels; L-Type; Cell-Penetrating Peptides; Cyclic AMP; Gene Expression; HEK293 Cells; Humans; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels; Membrane Proteins; Mice; Mice; Inbred C57BL; Molecular Docking Simulation; Myocytes; Cardiac; Patch-Clamp Techniques; Peptides; Peroxins; Potassium Channels; Protein Binding; Protein Conformation; alpha-Helical; Protein Conformation; beta-Strand; Protein Intera; Rabbits; Sinoatrial Node; tat Gene Products; Human Immunodeficiency Virus
Elenco autori:
Moroni, Anna
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