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Successive detection of insulin‐like growth factor‐II bound to receptors on a living cell surface using an AFM
Author(s) -
Han SungWoong,
Mieda Shingo,
Nakamura Chikashi,
Kihara Takanori,
Nakamura Noriyuki,
Miyake Jun
Publication year - 2009
Publication title -
journal of molecular recognition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.401
H-Index - 79
eISSN - 1099-1352
pISSN - 0952-3499
DOI - 10.1002/jmr.994
Subject(s) - atomic force microscopy , receptor , growth factor , insulin like growth factor , cell , biophysics , chemistry , surface (topology) , nanotechnology , materials science , biology , biochemistry , mathematics , geometry
In this study, we have developed a method of mechanical force detection for ligands bound to receptors on a cell surface, both of which are involved in a signal transduction pathway. This pathway is an autocrine pathway, involving the production of insulin‐like growth factor‐II (IGF‐II) and activation of the IGF‐I receptor, involved in myoblast differentiation induced by MyoD in C3H10T1/2 mouse mesenchymal stem cells. Differentiation of C3H10T1/2 was induced with the DNA demethylation agent 5‐azacytidine (5‐aza). The etched AFM tip used in the force detection had a flat surface of which about 10 µm 2 was in contact with a cell surface. The forces required to rupture the interactions of IGF‐IIs on a cell and anti mouse IGF‐II polyclonal antibody immobilized on an etched AFM tip were measured within 5 days of induction of differentiation. The mean unbinding force for a single paired antibody–ligand on a cell was about 81 pN, which was measured at a force loading rate of about 440 nN/s. The percentage of unbinding forces over 100 pN increased to 32% after 2 days from the addition of 5‐aza to the medium. This method could be used in non‐invasive and successive evaluation of a living cell's behavior. Copyright © 2009 John Wiley & Sons, Ltd.