A long acidic domain affects the chromatographic behaviour of a neuronal adaptor protein on DEAE-Sepharose.
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.
Longo
Olimpia
O
Department of Biochemistry and Medical Biotechnologies, University of Naples Federico II, Italy.
Lamberti
Annalisa
A
Zambrano
Nicola
N
Arcari
Paolo
P
D000818:Animals; D015153:Blotting, Western; D002122:Calcium Chloride / Q000737:chemistry / Q000378:metabolism; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods; D015636:Magnesium Chloride / Q000737:chemistry / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000378:metabolism; D009687:Nuclear Proteins / Q000737:chemistry* / Q000378:metabolism; D009994:Osmolar Concentration; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary; D051381:Rats; D012685:Sepharose / Q000031:analogs & derivatives* / Q000378:metabolism*
C486242:Apbb1 protein, rat; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D010446:Peptide Fragments; D015636:Magnesium Chloride; D012685:Sepharose; D002122:Calcium Chloride