Small-molecule mimics of an α-helix for efficient transport of proteins into cells

Abstract

We designed and synthesized small-molecule mimics of an alpha-helical peptide protein transduction domain (PTD). These small-molecule carriers, which we termed SMoCs, are easily coupled to biomolecules, and efficiently deliver dye molecules and recombinant proteins into a variety of cell types. We designed the SMoCs using molecular modeling techniques. As an example of a protein cargo, we applied this new technology to the internalization of the DNA replication licensing repressor geminin, in vitro, providing evidence that extracellularly delivered SMoC-geminin can have an antiproliferative effect on human cancer cells. Uptake of SMoC-geminin was inhibited at 4 degrees C and by chlorpromazine, a compound that induces misassembly of clathrin-coated pits at the cell surface. Thus the mechanism of uptake is likely to be clathrin-mediated endocytosis.

TRAIL induces endocytosis of its death receptors in MDA-MB-231 breast cancer cells

“…Treatment with chlorpromazine (CPZ), an inhibitor of clathrin-mediated endocytosis, 17,18 partially inhibited TRAIL-induced reduction of DR4 and DR5 from cell surface (Fig. 5B).…”

Genomics screen in transformed stem cells reveals RNASEH2A, PPAP2C, and ADARB1 as putative anticancer drug targets

“…6 We selected 146 samples hybridized to Affymetrix hgu133plus2 genechips, covering 11 common types of carcinoma (breast, colon, endometrium, kidney clearcell carcinoma, kidney papillary carcinoma, lung adenocarcinoma, ovary, prostate, rectum, thyroid, and bladder) with no fewer than 10 of each type. Our mesenchymal tumor expression data represent 19 different subtypes, including alveolar rhabdomyosarcoma, chondroblastoma, chondrosarcoma, chordoma, chondromyxoid fibroma, dedifferentiated chondrosarcoma, embryonal rhabdomyosarcoma, Ewing's sarcoma, fibromatosis, lipoma, leiomyosarcoma, myxoid liposarcoma, synovial sarcoma, malignant peripheral nerve sheath tumor, neurofibroma, osteosarcoma, pleiomorphic sarcoma, schwannoma, and well-differentiated liposarcoma (15).…”

“…5 expO repository (https://expo.intgen.org/geo/home.do). 6 National Center for Biotechnology Information Gene Expression Omnibus (ftp://ftp.ncbi.nlm.nih.gov/pub/geo/DATA/SeriesMatrix/GSE2109). 7 Oncomine (http://www.oncomine.org).…”

“…4). Recent advances, including antisense delivery, small interfering RNA (siRNA) knockdown, and small-molecule carriers, may extend the range of proteins that could be targeted (6,7).…”

Cell-Based GATA4 Cardiac Gene Transfer Using Cell-Penetrating Peptide

“…Developing small molecules which mimic CPP function may also help to increase trafficking efficiency. 31 (2) Targeting to specific cells. Resident cardiac fibroblasts contribute to the development and progression of CHF.…”