Work

Efficiency of Protein Transduction Is Cell Type-dependent and Is Enhanced by Dextran Sulfate

Year: 2002

Type: article

Abstract: Protein transduction domains (PTDs), both naturally occurring and synthetic, have been increasingly utilized to deliver biologically active agents to a variety of cell types in vitro and in vivo. We report that in addition to previously characterized arginine-rich PTDs, including TAT, lysine homopolymers were able to mediate transduction of a wide variety of cell types, as measured by flow cytometric and enzymatic assays. The efficiency of PTD-mediated transduction was influenced by the cell type tested, although polylysine homopolymers demonstrate levels of internalization that consistently exceeded those of TAT and arginine homopolymers. Transduction of arginine/lysine-rich PTDs occurred at 4 degrees C and following depletion of cellular ATP pools, albeit generally at reduced levels. Although transduction was reduced in Chinese hamster ovary mutant lines deficient in either heparan sulfate or glycosaminoglycan synthesis, uptake was restored to wild-type levels by incubating target cells with dextran sulfate. The enhancement of transduction by dextran sulfate suggests that electrostatic interactions play an important first step in the process by which PTDs and their cargo traverse the plasma membrane.

Source: Journal of Biological Chemistry

DOI: 10.1074/jbc.m204202200

Authors: Jeffrey C. Mai, Hongmei Shen, Simon C. Watkins, Tao Cheng, Paul D. Robbins

Institutions: University of Pittsburgh, UPMC Hillman Cancer Center

Language: English

Topic: RNA Interference and Gene Delivery

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: hybrid

Estimated APC paid: $3,430

Funder: National Institute of Arthritis and Musculoskeletal and Skin Diseases