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What can electron microscopy tell us about chaperoned protein folding?

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Presentation on theme: "What can electron microscopy tell us about chaperoned protein folding?"— Presentation transcript:

1 What can electron microscopy tell us about chaperoned protein folding?
Helen R Saibil  Folding and Design  Volume 1, Issue 3, Pages R45-R49 (June 1996) DOI: /S (96) Copyright © 1996 Elsevier Ltd Terms and Conditions

2 Figure 1 Plunger for vitrifying EM grids. The grid, with a small drop of solution, is held by forceps, blotted with filter paper to leave a thin film of liquid, and then immediately plunged into a coolant, where it solidifies to a glass-like state within ∼1 ms. If the plunger is coupled to the atomizer spray, a reactant can be sprayed onto the grid as fine droplets, just as the grid descends towards the coolant. The interval can be adjusted so that the spray is added a few milliseconds before vitrification. Diagram adapted from [7]. Folding and Design 1996 1, R45-R49DOI: ( /S (96) ) Copyright © 1996 Elsevier Ltd Terms and Conditions

3 Figure 2 Illustration of angular refinement for a GroEL–GroES complex. Top row, a set of surface-rendered views of a 3D reconstruction of the GroEL–GroES complex. From the left, the orientations are: sevenfold axis vertical; sevenfold axis vertical and 25° rotation around the sevenfold axis; sevenfold axis tipped forward 45°; and sevenfold axis perpendicular to the plane of view. Bottom row, 2D projections of the structure in the corresponding directions. Folding and Design 1996 1, R45-R49DOI: ( /S (96) ) Copyright © 1996 Elsevier Ltd Terms and Conditions

4 Figure 3 Schematic illustration of correspondence analysis, used to classify side views of GroEL–GroES complexes as full (containing folding substrate) or empty (no substrate). The top row represents a two-image data set, with image 1 containing substrate in the open binding site and image 2 devoid of substrate. Their average has half the substrate density. The eigenimage from correspondence analysis gives the difference density that must be added to or subtracted from the average image in order to regenerate images 1 and 2, respectively. Folding and Design 1996 1, R45-R49DOI: ( /S (96) ) Copyright © 1996 Elsevier Ltd Terms and Conditions


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