Single protein molecule mapping with magnetic atomic force microscopy.
Understanding the structural organization and distribution of proteins in biological cells is of fundamental importance in biomedical research. The use of conventional fluorescent microscopy for this purpose is limited due to its relatively low spatial resolution compared to the size of a single protein molecule. Atomic force microscopy (AFM), on the other hand, allows one to achieve single-protein resolution by scanning the cell surface using a specialized ligand-coated AFM tip. However, because this method relies on short-range interactions, it is limited to the detection of binding sites that are directly accessible to the AFM tip. We developed a method based on magnetic (long-range) interactions and applied it to investigate the structural organization and distribution of endothelin receptors on the surface of smooth muscle cells. Endothelin receptors were labeled with 50-nm superparamagnetic microbeads and then imaged with magnetic AFM. Considering its high spatial resolution and ability to "see" magnetically labeled proteins at a distance of up to 150 nm, this approach may become an important tool for investigating the dynamics of individual proteins both on the cell membrane and in the submembrane space.
Department of Physics, University of Bath, Bath, United Kingdom.
Andriy V
AV
Moskalenko
Polina L
PL
Yarova
Sergey N
SN
Gordeev
Sergey V
SV
Smirnov
D000818:Animals; D001011:Aorta / Q000166:cytology / Q000378:metabolism; D019921:Biotinylation; D002478:Cells, Cultured; D004574:Electromagnetic Fields; D019332:Endothelin-1 / Q000378:metabolism; D016650:Fluorescein-5-isothiocyanate; D021621:Imaging, Three-Dimensional / Q000379:methods; D008297:Male; D018625:Microscopy, Atomic Force / Q000379:methods*; D018613:Microscopy, Confocal; D008863:Microspheres; D032342:Muscle Cells / Q000166:cytology / Q000378:metabolism; D009130:Muscle, Smooth / Q000166:cytology / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar
D019332:Endothelin-1; D016650:Fluorescein-5-isothiocyanate