Problems regarding reconstitution

The fundamental problems regarding reconstitution of connexin channels have been overcome connexin forms channels in unilamellar liposomes and planar bilayers. Size, permeability, and gating behavior are consistent with conducting units that are single hemichannels—the structures that span a single cell membrane and form one-half of the junctional channel. Connexin can be obtained by affinity purification under nondenaturing conditions. Thus, channels formed by a single connexin can be studied in a well-defined and accessible system. 

Non-heme iron proteins frequently require reconstitution with iron after they are isolated. Reconstitution of P. furiosus SOR yields samples with iron in two kinds of site, each with a unique EPR spectrum. Determining which EPR spectrum corresponds to die catalytieally aetive iron site required eorrelation of an increase in one widi increases in enzyme activity. In addition, assignment of one of the EPR resonances to the redox active iron component was made by quantitative comparison of ferric signals obtained at points through a dye-mediated redox titration [8], In a similar problem regarding reconstituted phenylalanine hydroxylase (PAH),... 

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