Antibody-free lanthanide-based fluorescent probe for determination of protein tyrosine kinase and phosphatase activities

Autor: Aura Sipponen, Antti Valanne, Timo Lövgren, Pauliina Niemelä, Heidi Appelblom, Tero Soukka
Rok vydání: 2010
Předmět:
Zdroj: Microchimica Acta. 172:25-29
ISSN: 1436-5073
0026-3672
Popis: 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.
Databáze: OpenAIRE
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