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High speed AFM observation of sparse binding of individual cofilin-rod molecules to actin filaments.

National Institute of Advanced Industrial Science and Technology (AIST) and Kanazawa University Kien Xuan Ngo, Noriyuki Kodera Taro Q.P. Uyeda

Individual cofilin molecules are too small to visualize by AFM. Thus, we fused the rod domain of α-actinin to the C-terminus of cofilin (cofilin-rod), so that transient binding of cofilin-rod to and dissociation from an actin filament was visualized by the presence of a rod-like structure sticking out of the actin filament (blue arrowhead). Note that no severing was observed near singly bound cofilin-rod molecules. Conditions: F buffer containing 1 mM ATP and 75 nM cofilin-rod, imaging rate: 4 frames/s, and playing rate: 5 frames/s. The width of the imaged field: 150 nm, Z-scale: 0–12 nm.

eLife, 4:e04806, 2015

(2015.06.18)

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