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17
CONTENTS
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© 2002 by CRC Press LLC
Protein Transport
Jehangir S. Wadia, Michelle Becker–Hapak,
and Steven F. Dowdy
17.1 Protein Transduction ....................................................................................365
17.2 Protein Transduction Technology ................................................................366
17.3 Development of PTD Fusion Proteins.........................................................367
17.3.1 Cloning and Purification of Tat Fusion Proteins.............................368
17.4 Delivery of PTD-Conjugated Macromolecules ...........................................369
17.4.1 Applications in Cell Culture............................................................369
17.4.2 In Vivo Applications.........................................................................371
17.5 Future Directions and Considerations .........................................................373
References..............................................................................................................373
17.1 PROTEIN TRANSDUCTION
The average eukaryotic cell contains more than 10,000 different proteins that participate
in normal cellular functions such as signal transduction, gene transcription,
cell-to-cell communication, and protein trafficking. When deregulated, these are
often involved in the onset and progression of disease. Importantly, proteins have
been evolutionarily selected to perform very specific functions. Thus, the ability to
manipulate the cellular level of these proteins or the ectopic expression of novel
proteins has shown tremendous potential as a biological tool for studying cellular
processes and may become a cornerstone in the effective treatment of human disease.
For instance, the reconstitution of tumor-suppressor function by the introduction of
tumor-suppressor proteins in cancer therapy or the replacement of defective proteins
in genetic disease such as cystic fibrosis or Duchenne’s muscular dystrophy are often
considered the goal of effective treatment. 1
In practice, however, the ideal pharmacological agents to achieve these goals
and their associated delivery must overcome several key obstacles, including poor
specificity, bioavailability, tissue distribution, and toxicity. These obstacles currently
limit the usefulness of techniques such as drug therapy or genetic approaches and
largely account for failure of the early promise afforded by introduction of recombinant
protein technology. For instance, over the past century, the serendipitous
discovery of certain drugs and small molecules which could be delivered easily and
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