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Published on: March 10, 2010
A long acidic domain affects the chromatographic behaviour of a neuronal adaptor protein on DEAE-Sepharose
Olimpia Longo1, Annalisa Lamberti, Nicola Zambrano
1Department of Biochemistry and Medical Biotechnologies, University of Naples Federico II, Italy.
Abstract:
The stepwise chromatographic behaviour on DEAE-Sepharose of rat Fe65, a neuronal protein, was tested, using as eluants KCl, CaCl2, and MgCl2. Assays by western blot showed that Fe65 was eluted by CaCl2, at a ionic strength 20% lower than that of MgCl2 or KCl. Interestingly, in the case of a truncated Fe65, lacking a glutamic acid rich region at the N-terminus, the ionic strengths of the various eluants were almost identical. These results suggested a possible inhibitory role of calcium ions in the binding of the protein to DEAE and a specific affinity of these ions for long acidic stretches.
