Coupling to polymeric scaffolds stabilizes biofunctional peptides for intracellular applications

I. Ruttekolk, A. Chakrabarti, M. Richter, F. Duchardt, H. Glauner, W.P.R. Verdurmen, J. Rademann, R. Brock – 2011

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.

Titel
Coupling to polymeric scaffolds stabilizes biofunctional peptides for intracellular applications
Verfasser
I. Ruttekolk, A. Chakrabarti, M. Richter, F. Duchardt, H. Glauner, W.P.R. Verdurmen, J. Rademann, R. Brock
Datum
2011
Kennung
10.1124/mol.110.068296
Zitierweise
Mol. Pharmac. 2011, 79(4), 692-700
Art
Text
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