Abstract
AFM is a powerful technique for revealing the morphological features of various biological systems at high resolution. However, one of the complications of AFM is that samples must be attached to a flat surface in order to obtain images. This often requires the development of specialized methods depending on the sample which is being used. In this study, we developed a novel technique to image actin bundles on the mica surface. Using this technique, we were able to image molecular assemblies of F-actin with two actin remodeling proteins: α-actinin and Caprice. High resolution AFM images of F-actin fibers and bundle organization depicted two different types of molecular assemblies: F-actin bundles forming an elongated “zipper” structure in the presence of α-actinin, and bundles forming a perpendicularly crossing the mesh structure in the presence of Caprice.
Highlights
The biological applications of atomic force microscopy (AFM) [1] were realized immediately after the instrument was invented in 1986 [2]
The samples were prepared using a typical deposition procedure in which either 10 μL 900 ng/μL filamentous actin (F-actin) or F-actin/Caprice was deposited on freshly cleaved mica, washed with 3 mL of water, and dried by blowing with a stream of nitrogen gas
We have developed a strategy to image actin bundles with AFM by centrifuging the preformed bundles onto a mica surface
Summary
The biological applications of atomic force microscopy (AFM) [1] were realized immediately after the instrument was invented in 1986 [2]. The physical properties of DNA, proteins, lipids and carbohydrates have been extensively studied by using AFM [3,4,5,6]. The development of new scanning methods [7] and cantilever production [8] has facilitated the applications of AFM for studying biological macromolecules in physiological conditions. The most fascinating technical endeavor has been the invention of high-speed AFM by Ando et al, which has enabled one to capture the motion of DNA [9,10,11,12] and proteins [13,14,15,16,17] and to monitor enzymatic reactions [9,11, 18] in solution
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