Signal mutagenesis

Labelling Na,K-ATPase with ATP analogues provides evidence for contribution from charged residues that are widely separated in the sequence of a subunit of Na,K-ATPase. The first indication came from ATP sensitive covalent insertion of fluorescein-isothiocyanate (FITC) into Lys ° in the a subunit [90], The strong fluorescence signal provides a convenient probe for monitoring conformational transitions in the proteins. Site-directed mutagenesis of Lys reduces the activity of... 

Since these proteins are difficult to crystallize and site-directed mutagenesis has certain limitations, PAL can find much use in exploring pathways, transducers, and effectors in signal transduction as well as some of their main structural features. 

Niviere, V., Wong, S. L. and Voordouw, G. (1992) Site-directed mutagenesis of the hydro-genase signal peptide consensus box prevents export of a beta-lactamase fusion protein. J. Gen. Microbiol., 138, 2173-83. 

Kristiansen K. Molecular mechanisms of ligand binding, signalling, and regulation within the superfamily of G-protein-coupled receptors molecular modeling and mutagenesis approaches to receptor structure and function. Pharmacol Ther 2004 103 21-80. 

The inducible systems described above, apart from utilizing them for the induction of EOF mutations through gene deletion (Fig. 2), can also be exploited for the inducible expression of cDNAs or shRNAs. This is achieved by crossing mice that have a transcriptional STOP signal, flanked by loxP sites, between the promoter and the GOI cDNA or the shRNA (Fig. lb, e) to one of the tool mice for conditional mutagenesis (Fig. 2b-d). 

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