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A TAT-streptavidin fusion protein directs uptake of biotinylated cargo into mammalian cells


, : A TAT-streptavidin fusion protein directs uptake of biotinylated cargo into mammalian cells. Protein Engineering, Design & Selection 18(3): 147-152

The HIV-1 TAT peptide has been used extensively for directing the intracellular delivery of an assortment of cargo, including DNA, liposomes and macromolecules. For protein delivery, a variety of TAT-fusion proteins have been described which link the TAT coding sequence to the protein coding sequence of interest. Streptavidin represents a potentially useful TAT-fusion protein because it could be used to deliver a wide array of biotinylated cargo. Here we have characterized a TAT-streptavidin (TAT-SA) fusion protein, which retains the ability to bind biotinylated cargo while directing their efficient cellular uptake. Fluorescence activated cell sorting (FACS) analysis and confocal microscopy characterization showed that TAT-SA is internalized by Jurkat T-cells and NIH 3T3 cells alone and when complexed to phycoerythrin, whereas the native streptavidin is not. Additionally, biotinylated alkaline phosphatase is successfully internalized and retains its activity when complexed to TAT-SA and incubated with Jurkat T-cells. Confocal microscopy suggested, however, that internalized TAT-SA and TAT-SA complexes were largely compartmentalized in vesicular compartments, rather than freely diffusing in the cytoplasmic compartment. To effect cytoplasmic delivery, the endosomal releasing polymer, poly(propylacrylic acid) (PPAA), was biotinylated and complexed to TAT-SA. Endosomal release and cytoplasmic delivery of fluorescently labeled TAT-SA complexes with PPAA was shown by the diffuse distribution of fluorescent protein in the cytoplasm. Taken together, these results demonstrate that TAT-SA can be used to direct intracellular delivery of large biotinylated cargo to intracellular compartments and that biotinylated PPAA can direct cytoplasmic delivery where desired.

US$19.90

PMID: 15820981

DOI: 10.1093/protein/gzi014


Other references

Ferry, G.D.; Büller, H.A., 1995: Mechanisms of growth retardation, drug therapy, and nutritional support in pediatric inflammatory bowel disease: a workshop sponsored by the north american and European societies for pediatric gastroenterology and nutrition. : Growth retardation is common in children with inflammatory bowel disease (IBD). The most sensitive measure of impaired growth, growth velocity, is abnormal in 65% of children with Crohn's disease. With treatment, growth velocity may return...

Fuentes, B.; Díez-Tejedor, E., 2000: The advantages of a stroke unit in the treatment of intracerebral hemorrhage. The efficacy of a stroke unit in the management of acute cerebrovascular disease has, in general, been widely demonstrated. Our analysis seeks to determine whether this efficacy is maintained in intracerebral hemorrhage. We studied the patients wi...

Masters, B.R.; Fanestil, D.D., 1979: Metabolic dependence of the offset of antidiuretic hormone-induced osmotic flow of water across the toad urinary bladder. The elevated osmotic permeability to water induced by antidiuretic hormone (ADH) in the isolated urinary bladder of the toad is rapidly reversed by removal or washout of the ADH. This return to normal water permeability is delayed by the suppressi...

Gaviria-Agudelo, C.L.; Jordan-Villegas, A.; Garcia, C.; McCracken, G.H., 2016: The Effect of 13-Valent Pneumococcal Conjugate Vaccine on the Serotype Distribution and Antibiotic Resistance Profiles in Children With Invasive Pneumococcal Disease. Invasive pneumococcal disease (IPD) continues to be a significant burden in children despite the implementation of two generations of conjugate vaccines. Serotype replacement by nonvaccine serotypes is reported in multiple areas around the world....

Reynolds, C., 2004: The Freshwater Biological Association at 75. Freshwater Forum, 22: 3-12

Anderson, R.A., 1941: Fusulinids of the Granite Falls limestone and their stratigraphic significance. Res Stud State College Washington, 9(3): 189-202

Pax, Ferdinand., 1907: Vorarbeiten zu einer Revision der Familie Actiniidae. (Diss.). Unknown

Loudon, J.A., 2013: Ataluren: a 'no-nonsense' approach for pulmonary diseases. Pulmonary Pharmacology & Therapeutics 26(3): 398-399

Bourne, D.G.; Holmes, A.J.; Iversen, N.; Murrell, J.C.lin, 2000: Fluorescent oligonucleotide rDNA probes for specific detection of methane oxidising bacteria. Oligonucleotide probes targeting the 16S rRNA of distinct phylogenetic groups of methanotrophs were designed for the in situ detection of these organisms. A probe, MG-64, detected specifically type I methanotrophs, while probes MA-221 and MA-621,...

Kusakari, J.; Ohyama, K.; Inamura, N.; Ikeda, K.; Arakawa, E.; Kaneko, Y.; Takasaka, T.; Kawamoto, K., 1985: Gas analysis of the middle ear cavity in normal and pathological conditions. The gas of the tympanic bulla was measured using 18 normal and 14 tube-obstructed guinea pigs. The concentrations of CO2 and O2 were 4.8 +/- 1.0% and 7.2 +/- 1.8% in normal animals, and 4.3 +/- 1.6% and 8.1 +/- 2.0% in tube-obstructed animals, res...