Fluorescent protein basics

Fluorescence spectroscopy, of proteins basic theory and interpretation, 257-263 determination of fluorescence quenching, 253-255... 

Ever since the demonstration that synthetic peptides could target a protein to the nucleus when cross-linked to it (Goldfarb et ai, 1986), peptide-protein conjugates have been used widely in nuclear protein import assays in vivo and in vitro. The method described here is for the preparation of a fluorescent bovine serum albumin, SV40 NLS conjugate. However, this basic protocol can be adapted easily to allow the conjugation to other proteins, such as the naturally fluorescent protein allophycocyanin (Adam etai, 1991). 

The conformation of bovine myelin basic protein (MBP) in AOT/isooctane/water reversed micellar systems was studied by Waks et al. 67). This MBP is an extrinsic water soluble protein which attains an extended conformation in aqueous solution 68 but is more density packed at the membrane surface. The solubilization of MBP in the AOT reversed micelles depends on the water/AOT-ratio w0 68). The maximum of solubilization was observed at a w0-value as low as 5.56. The same value was obtained for another major protein component of myelin, the Folch-Pi proteolipid 69). According to fluorescence emission spectra of MBP, accessibility of the single tryptophane residue seems to be decreased in AOT reversed micelles. From CD-spectra one can conclude that there is a higher conformational rigidity in reversed micelles and a more ordered aqueous environment. 

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