caa a22 4500 445879904 CHVBK 20180317145539.0 cr unu---uuuuu 170323e20110201xx s 000 0 eng 10.1007/s00604-010-0450-x doi (NATIONALLICENCE)springer-10.1007/s00604-010-0450-x Antibody-free lanthanide-based fluorescent probe for determination of protein tyrosine kinase and phosphatase activities [Elektronische Daten] [Heidi Appelblom, Aura Sipponen, Antti Valanne, Tero Soukka, Timo Lövgren, Pauliina Niemelä] A single-labeled peptide probe for measuring peptide phosphorylation status was developed by using a phosphate sensitive terbium chelate. The activity of Abl protein tyrosine kinase and T-cell protein Tyrosine phosphatase (TC PTP) was monitored in real time. To study the probe design in detail, variable substrate peptide sequences, where the enzyme target site was located from two to five amino acids apart from the nearest tyrosine residue, were synthesized. The maximum change observed in fluorescence intensity after phosphorylation was up to 320%, when the phosphorylated tyrosine was located two amino acids from the lysine coupled to the phosphate sensitive terbium chelate, demonstrating an excellent performance for a homogeneous assay. Also the longer distance of five amino acids between the phosphorylated tyrosine residue and terbium chelate resulted up to 260% change in fluorescence intensity. Figure A principle of the short peptide probe (EAI‐Y‐AAPFAK) with phosphate sensitive terbium chelate attached to the lysine side chain is described, which is proved applicable to measure in real time Abl protein tyrosine kinase and T‐cell protein tyrosine phosphatase activities. Enhancement of the terbium fluorescence could be measured upon addition of a phosphor residue to the nearby tyrosine side chain. The opposite effect could be measured, when phosphor residue is removed by protein tyrosine phosphatase. Springer-Verlag, 2010 Protein tyrosine kinase nationallicence Protein tyrosine phosphatase nationallicence Biosensor nationallicence Time-resolved fluorescence nationallicence Lanthanide chelate nationallicence Appelblom Heidi Department of Biotechnology, University of Turku, Tykistökatu 6A, 20520, Turku, Finland aut Sipponen Aura Department of Biotechnology, University of Turku, Tykistökatu 6A, 20520, Turku, Finland aut Valanne Antti Department of Biotechnology, University of Turku, Tykistökatu 6A, 20520, Turku, Finland aut Soukka Tero Department of Biotechnology, University of Turku, Tykistökatu 6A, 20520, Turku, Finland aut Lövgren Timo Department of Biotechnology, University of Turku, Tykistökatu 6A, 20520, Turku, Finland aut Niemelä Pauliina Department of Biotechnology, University of Turku, Tykistökatu 6A, 20520, Turku, Finland aut Microchimica Acta Springer Vienna 172/1-2(2011-02-01), 25-29 0026-3672 172:1-2<25 2011 172 604 https://doi.org/10.1007/s00604-010-0450-x text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00604-010-0450-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Appelblom Heidi Department of Biotechnology, University of Turku, Tykistökatu 6A, 20520, Turku, Finland aut NATIONALLICENCE 700 1- Sipponen Aura Department of Biotechnology, University of Turku, Tykistökatu 6A, 20520, Turku, Finland aut NATIONALLICENCE 700 1- Valanne Antti Department of Biotechnology, University of Turku, Tykistökatu 6A, 20520, Turku, Finland aut NATIONALLICENCE 700 1- Soukka Tero Department of Biotechnology, University of Turku, Tykistökatu 6A, 20520, Turku, Finland aut NATIONALLICENCE 700 1- Lövgren Timo Department of Biotechnology, University of Turku, Tykistökatu 6A, 20520, Turku, Finland aut NATIONALLICENCE 700 1- Niemelä Pauliina Department of Biotechnology, University of Turku, Tykistökatu 6A, 20520, Turku, Finland aut NATIONALLICENCE 773 0- Microchimica Acta Springer Vienna 172/1-2(2011-02-01), 25-29 0026-3672 172:1-2<25 2011 172 604 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer