Abstract
Obtaining structural information from single biomolecules using imaging methods is challenging due to resolution limitations and difficulties in labelling specific sites of biomolecules. Atomic force microscopy based imaging of interaction forces between a specially designed probe molecule and a biomolecular complex of interest can, in principle, allow determining the locations of various regions within the complex, if the probe is designed to exhibit affinity to these regions. Here we demonstrate this concept with protein-RNA complex by targeting specific segments of the RNA with a DNA probe having sequence complementarity with these segments. To map the interaction forces between the probe and the sample, we used T-shaped cantilevers that allow mapping interaction forces during the tapping mode imaging process [1]. We worked with stem loop binding proteins in complex with RNA [2] and we targeted the flanking RNA regions adjacent to the stem loop. We show that locations and sequence identities of the RNA segments can be imaged with this approach. Repeated measurements of the same protein-RNA complex showed the consistency of the images. Our results show that it is possible obtain chemically-specific structural information from single biomolecules in physiologically relevant conditions without using labels.1. Kim, D., Sahin, O. Nature Nanotechnology 10, 264-269 (2015).2. Tan, D., Mazluff, W. F., Dominski, Z., Tong, L. Science 339, 318-321 (2013).
Published Version
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