ISSN 2410-7751 (Print)
ISSN 2410-776X (Online)
"Biotechnologia Acta" v. 7, no 2, 2014
Р. 46-53, Bibliography 11, Ukrainian.
Universal Decimal classification: 577(112+25+354.9)+57(085.2+086.2)
https://doi.org/10.15407/biotech7.02.046
OBTAINING AND CHARACTERIZATION OF RECOMBINANT FLUORESCENT DERIVATIVES OF SOLUBLE HUMAN HB-EGF
N. V. Korotkevich, A. Ju. Labyntsev, D. V. Kolibo, S. V. Komisarenko
Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, Kyiv
Heparin-binding EGF-like growth factor (HB-EGF) belongs to the epidermal growth factor receptor family and is synthesized as a transmembrane precursor proHB-EGF. Binding of sHB-EGF with receptors EGFR and HER-4 results in formation of ligand-receptor complexes and activation of signaling pathways and thereby promotes survival, proliferation and migration of cells.
The aim of the study was to obtain the recombinant fluorescent derivatives of full-length soluble sHB-EGF – mCherry-sHB-EGF and truncated sHB-EGF?84-106 (without heparin-binding domain) – mCherry-sHB-EGF?84-106. The recombinant fluorescent derivatives may be used to investigate the sHB-EGF binding with receptors EGFR and HER-4, intracellular transportation of ligand-receptor complexes and the role of sHB-EGF heparin-binding domain in sHB-EGF biological activity.
It was shown the ability of both fluorescent derivatives specifically bind to EGFR, to internalize in receptor-mediated pathway and to enhance the proliferation of 3T3 cells. Absence of heparin-binding domain in structure of mCherry-sHB-EGF?84-106 significantly did not effect on its ability to bind with receptor, but decreased twice its mitogenic activity. Thus, obtained fluorescent derivatives of sHB-EGF could be a convenient tool for investigation of molecular mechanisms of sHB-EGF biological activity and role of heparin-binding domain in these processes.
Key words: heparin-binding EGF-like growth factor (HB-EGF), epidermal growth factor receptor.
© Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, 2014
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