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  • Research Article
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Genetic engineering of proteins with cell membrane permeability

Nature Biotechnology volume 16, pages 370–375 (1998)Cite this article

Abstract

The discovery of methods for generating proteins with inherent cell membrane-translocating activity will expand our ability to study and manipulate various intracellular processes in living systems. We report a method to engineer proteins with cell-membrane permeability. After a 12–amino acid residue membrane-translocating sequence (MTS) was fused to the C-terminus of glutathione S-transferase (GST), the resultant GST-MTS fusion proteins were efficiently imported into NIH 3T3 fibroblasts and other cells. To explore the applicability of this nondestructive import method to the study of intracellular processes, a 41-kDa GST-Grb2SH2-MTS fusion protein containing the Grb2 SH2 domain was tested for its effect on the epidermal growth factor (EGF)-stimulated signaling pathway. This fusion protein entered cells, formed a complex with phosphorylated EGF receptor (EGFR), and inhibited EGF-induced EGFR–Grb2 association and mitogen-activated protein kinase activation.

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Authors and Affiliations

  1. Department of Microbiology and Immunology, Vanderbilt University, Nashville, TN, 37232

    Mauricio Rojas, Zhongjia Tan & Yao-Zhong Lin

  2. Division of Infectious Diseases, Department of Medicine, Vanderbilt University, Nashville, TN, 37232

    John P. Donahue

Authors
  1. Mauricio Rojas
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  2. John P. Donahue
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  3. Zhongjia Tan
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  4. Yao-Zhong Lin
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Correspondence to Yao-Zhong Lin.

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Rojas, M., Donahue, J., Tan, Z. et al. Genetic engineering of proteins with cell membrane permeability. Nat Biotechnol 16, 370–375 (1998). https://doi.org/10.1038/nbt0498-370

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  • DOI: https://doi.org/10.1038/nbt0498-370

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