Controllable protein cleavages through intein fragment complementation
نویسندگان
چکیده
منابع مشابه
Protein fragment complementation in M.HhaI DNA methyltransferase.
The 5mC DNA methyltransferase M.HhaI can be split into two individually inactive N- and C-terminal fragments that together can form an active enzyme in vivo capable of efficiently methylating DNA. This active fragment pair was identified by creating libraries of M.HhaI gene fragment pairs and then selecting for the pairs that code for an active 5mC methyltransferase. The site of bisection for s...
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Inteins are protein splicing elements that mediate their excision from precursor proteins and the joining of the flanking protein sequences (exteins). In this study, protein splicing was controlled by splitting precursor proteins within the Psp Pol-1 intein and expressing the resultant fragments in separate hosts. Reconstitution of an active intein was achieved by in vitro assembly of precursor...
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The organization of biochemical networks that make up the living cell can be defined by studying the dynamics of protein-protein interactions. To this end, experimental strategies based on protein fragment complementation assays (PCAs) have been used to map biochemical networks and to identify novel components of these networks. Pharmacological perturbations of the interactions can be observed,...
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We have developed a general experimental strategy that enables the quantitative detection of dynamic protein-protein interactions in intact living cells, based on protein-fragment complementation assays (PCAs). In this method, protein-protein interactions are coupled to refolding of enzymes from cognate fragments where reconstitution of enzyme activity acts as the detector of a protein interact...
متن کاملConstruction of protein fragment complementation libraries using incremental truncation.
Many proteins can have their peptide backbone cut by proteolytic or genetic means, yet the two fragments can associate to make an active heterodimer. This ‘‘monomer-to-heterodimer conversion’’ is referred to as protein fragment complementation (PFC). Such complementation is the reverse of evolutionary processes in which domains are recruited and fused at the genetic level. Classic examples of p...
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ژورنال
عنوان ژورنال: Protein Science
سال: 2009
ISSN: 0961-8368,1469-896X
DOI: 10.1002/pro.249