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A potential pitfall in protein kinase assay: phosphocellulose paper as un unreliable adsorbent of produced phosphopeptides
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical and Physiological Chemistry. (ek pia)
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical and Physiological Chemistry. (ek pia)
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical and Physiological Chemistry. (engström lorentz)
1992 (English)In: Analytical Biochemistry, ISSN 0003-2697, E-ISSN 1096-0309, Vol. 204, no 2, 311-314 p.Article in journal (Refereed) Published
Abstract [en]

The ability of phosphocellulose paper to retain phosphorylated peptides containing basic amino acid residues was investigated. Some peptide substrates that are commonly used for three different protein kinases were tested. The adsorption onto phosphocellulose paper was strongly dependent on the amino acid composition of the peptides. None of the phosphopeptides studied was adsorbed completely, the amount bound varied from 7 to 93%. Phosphopeptides containing two basic amino acids each differed remarkably in the degree of binding to the phosphocellulose paper (40% RRASVA, 60% FRRLSI, and 80% HRASV was bound). The results presented here indicate that data from phosphorylation experiments obtained so far for different peptides using the phosphocellulose paper method should be judged with caution.

Place, publisher, year, edition, pages
1992. Vol. 204, no 2, 311-314 p.
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Biochemistry and Molecular Biology
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URN: urn:nbn:se:uu:diva-173185DOI: 10.1016/0003-2697(92)90244-2OAI: oai:DiVA.org:uu-173185DiVA: diva2:516835
Available from: 2012-04-20 Created: 2012-04-20 Last updated: 2017-12-07Bibliographically approved

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