Methods of Analysis for Protein Dynamics in Living Cells Based on Protein Splicing

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Author(s)

Abstract

Protein–protein interactions and protein localization have key roles in many essential biological processes in living cells. We have developed novel reporter proteins with general applicability for detecting the biological processes in living cells and animals. The principle is based on reconstitution of split reporter proteins by protein splicing, which involves a self-catalyzed excision of a protein splicing element, intein, from flanking polypeptide sequences, exteins, leading to ligation of the flanking exteins by a peptide bond. As the exteins, N- and C-terminal halves of rationally-split green fluorescent protein (GFP) or split bioluminescent proteins were used. The N- and C-terminal reporters connected respectively with a pair of interacting proteins worked as indicators for protein–protein interactions in bacteria and mammalian cells. The split GFP reporter provided a genetic method for identifying mitochondrial proteins from large-scale complementary DNA libraries. The split bioluminescent reporter enabled high-throughput sensing and noninvasive imaging of nuclear transport of target proteins in living animals. This basic concept of split reporter reconstitution by protein splicing provides a wide variety of applications not only for fundamental biological studies, but also for assay and screening methods for chemicals that inhibit or facilitate the biological processes in living cells.

Journal

  • Bulletin of the Chemical Society of Japan

    Bulletin of the Chemical Society of Japan 78(5), 739-751, 2005-05-15

    The Chemical Society of Japan

References:  38

Codes

  • NII Article ID (NAID)
    10015716238
  • NII NACSIS-CAT ID (NCID)
    AA00580132
  • Text Lang
    ENG
  • Article Type
    REV
  • ISSN
    00092673
  • NDL Article ID
    7309400
  • NDL Source Classification
    ZP1(科学技術--化学・化学工業)
  • NDL Call No.
    Z53-B35
  • Data Source
    CJP  NDL  J-STAGE 
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