RT Journal Article
SR Electronic
T1 Coupling to Polymeric Scaffolds Stabilizes Biofunctional Peptides for Intracellular Applications
JF Molecular Pharmacology
JO Mol Pharmacol
FD American Society for Pharmacology and Experimental Therapeutics
SP 692
OP 700
DO 10.1124/mol.110.068296
VO 79
IS 4
A1 Ivo R. Ruttekolk
A1 Alokta Chakrabarti
A1 Martin Richter
A1 Falk Duchardt
A1 Heike Glauner
A1 Wouter P. R. Verdurmen
A1 Jörg Rademann
A1 Roland Brock
YR 2011
UL http://molpharm.aspetjournals.org/content/79/4/692.abstract
AB Here, we demonstrate that coupling to N-hydroxypropyl methacrylamide (HPMA) copolymer greatly enhances the activity of apoptosis-inducing peptides inside cells. Peptides corresponding to the BH3 domain of Bid were coupled to a thioester-activated HPMA (28.5 kDa) via native chemical ligation in a simple one-pot synthesis. Peptides and polymer conjugates were introduced into cells either by electroporation or by conjugation to the cell-penetrating peptide nona-arginine. The molecular basis of the increased activity is elucidated in detail. Loading efficiency and intracellular residence time were assessed by confocal microscopy. Fluorescence correlation spectroscopy was used as a separation-free analytical technique to determine proteolytic degradation in crude cell lysates. HPMA conjugation strongly increased the half-life of the peptides in crude cell lysates and inside cells, revealing proteolytic protection as the basis for higher activity.