Abstract
Proteins are the most critical executive molecules by responding to the instructions stored in the genetic materials in any form of life. More frequently, proteins do their jobs by acting as a roleplayer that interacts with other protein(s), which is more evident when the function of a protein is examined in the real context of a cell. Identifying the interactions between (or amongst) proteins is very crucial for the biochemistry investigation of an individual protein and for the attempts aiming to draw a holo-picture for the interacting members at the scale of proteomics (or protein-protein interactions mapping). Here, we introduced the currently available reporting systems that can be used to probe the interaction between candidate protein pairs based on the fragment complementation of some particular proteins. Emphasis was put on the principles and details of experimental design. These systems are dihydrofolate reductase (DHFR), β-lactamase, tobacco etch virus (TEV) protease, luciferase, β- galactosidase, GAL4, horseradish peroxidase (HRP), focal adhesion kinase (FAK), green fluorescent protein (GFP), and ubiquitin.
Keywords:
Protein-protein interaction; enzyme; fluorescent protein; interaction reporting system; protein complementation assay; ubiquitin.
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MeSH terms
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Animals
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Binding Sites
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Biological Assay*
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DNA-Binding Proteins / chemistry
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DNA-Binding Proteins / metabolism
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Endopeptidases / chemistry
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Endopeptidases / metabolism
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Escherichia coli / enzymology
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Focal Adhesion Protein-Tyrosine Kinases / chemistry
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Focal Adhesion Protein-Tyrosine Kinases / metabolism
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Green Fluorescent Proteins / chemistry
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Green Fluorescent Proteins / metabolism
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Horseradish Peroxidase / chemistry
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Horseradish Peroxidase / metabolism
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Luciferases / chemistry
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Luciferases / metabolism
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Peptide Fragments / analysis*
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Potyvirus / enzymology
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Protein Binding
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Protein Interaction Mapping / methods*
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Saccharomyces cerevisiae / enzymology
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Saccharomyces cerevisiae Proteins / chemistry
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Saccharomyces cerevisiae Proteins / metabolism
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Tetrahydrofolate Dehydrogenase / chemistry
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Tetrahydrofolate Dehydrogenase / metabolism
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Transcription Factors / chemistry
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Transcription Factors / metabolism
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Ubiquitin / chemistry
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Ubiquitin / metabolism
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beta-Galactosidase / chemistry
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beta-Galactosidase / metabolism
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beta-Lactamases / chemistry
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beta-Lactamases / metabolism
Substances
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DNA-Binding Proteins
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GAL4 protein, S cerevisiae
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Peptide Fragments
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Saccharomyces cerevisiae Proteins
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Transcription Factors
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Ubiquitin
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Green Fluorescent Proteins
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Horseradish Peroxidase
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Luciferases
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Tetrahydrofolate Dehydrogenase
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Focal Adhesion Protein-Tyrosine Kinases
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beta-Galactosidase
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Endopeptidases
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TEV protease
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beta-Lactamases