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American Society for Pharmacology and Experimental Therapeutics (ASPET), Molecular Pharmacology, 4(79), p. 692-700, 2011

DOI: 10.1124/mol.110.068296

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Coupling to polymeric scaffolds stabilizes biofunctional peptides for intracellular applications.

This paper is made freely available by the publisher.
This paper is made freely available by the publisher.

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Abstract

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.