Split-protein systems

Beyond binary protein-protein interactions

Sujan S. Shekhawat, Indraneel Ghosh

Research output: Contribution to journalArticle

82 Citations (Scopus)

Abstract

It has been estimated that 650,000 protein-protein interactions exist in the human interactome (Stumpf et al., 2008 [1]), a subset of all possible macromolecular partnerships that dictate life. Thus there is a continued need for the development of sensitive and user-friendly methods for cataloguing biomacromolecules in complex environments and for detecting their interactions, modifications, and cellular location. Such methods also allow for establishing differences in the interactome between a normal and diseased cellular state and for quantifying the outcome of therapeutic intervention. A promising approach for deconvoluting the role of macromolecular partnerships is split-protein reassembly, also called protein fragment complementation. This approach relies on the appropriate fragmentation of protein reporters, such as the green fluorescent protein or firefly luciferase, which when attached to possible interacting partners can reassemble and regain function, thereby confirming the partnership. Split-protein methods have been effectively utilized for detecting protein-protein interactions in cell-free systems, Escherichia coli, yeast, mammalian cells, plants, and live animals. Herein, we present recent advances in engineering split-protein systems that allow for the rapid detection of ternary protein complexes, small molecule inhibitors, as well as a variety of macromolecules including nucleic acids, poly(ADP) ribose, and iron sulfur clusters. We also present advances that combine split-protein systems with chemical inducers of dimerization strategies that allow for regulating the activity of orthogonal split-proteases as well as aid in identifying enzyme inhibitors. Finally, we discuss autoinhibition strategies leading to turn-on sensors as well as future directions in split-protein methodology including possible therapeutic approaches.

Original languageEnglish (US)
Pages (from-to)790-797
Number of pages8
JournalCurrent Opinion in Chemical Biology
Volume15
Issue number6
DOIs
StatePublished - Dec 2011

Fingerprint

Proteins
Ternary Complex Factors
Cataloging
Firefly Luciferases
Protein Engineering
Ribose
Cell-Free System
Regain
Plant Cells
Dimerization
Enzyme Inhibitors
Green Fluorescent Proteins
Sulfur
Nucleic Acids
Macromolecules
Yeast
Escherichia coli
Peptide Hydrolases
Iron
Yeasts

ASJC Scopus subject areas

  • Biochemistry
  • Analytical Chemistry

Cite this

Split-protein systems : Beyond binary protein-protein interactions. / Shekhawat, Sujan S.; Ghosh, Indraneel.

In: Current Opinion in Chemical Biology, Vol. 15, No. 6, 12.2011, p. 790-797.

Research output: Contribution to journalArticle

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