Mapping Protein Proteins Of A Living Cell

1424 WordsNov 5, 20146 Pages
Summary: Mapping protein-protein interactions occurring in a living cell can provide answers to plethora of questions in biological research. Split Beta-lactamase based protein fragment complementation assay provides an opportunity to analyze such interactions in both in vitro and in vivo environments. This flexible and robust assay holds potential to create a difference in our understanding of cellular processes. Introduction Protein-protein interactions form the basis of cellular processes that occur inside a living cell. Recent advances in biological research have strongly instigated the need to map biochemical networks for improved understanding of living cells. Apart from prediction and determination of protein structures to ascribe…show more content…
Protein fragment Complementation: What’s in the name? Protein fragment Complementation assays (PCAs) have emerged as an exciting option for such applications and tend to offer complete solution to study protein-protein interactions in a high-throughput format. These assays comprise of a protein molecule, which is rationally split into two fragments that do not interact with each other independently but can refold and reconstitute the activity of the native protein when brought together by interacting partners fused to them covalently. The reconstituted activity thus obtained can be used to select for such interacting pairs (Figure 1). To name a few, PCAs employing proteins like murine Dihydrofolate reductase (mDHFR), TEM-1 Beta lactamase (TEM-1 Bla), luciferase, Beta galactosidase, GFP, g3p of M13 filamentous phage have been used successfully for studying protein- protein interactions in bacterial hosts. TEM -1 Beta lactamase: The molecule of choice The focus of this review is the use of one of the most celebrated molecule the history of biology “TEM-1 Beta lactamase” (TEM-1 Bla; EC: 3.5.2.6) in PCAs. TEM-1 Bla is an E.coli derived plasmid borne resistance determinant to Beta lactam based drugs which was discovered in 1965 and named after the patient Temorina suffering from urinary tract infection untreatable with ampicillin. Although, clinically it has remained a barrier in the
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