αB-R120G-crystallin has more amyloid-like secondary structure than wild-type αB-crystallin proteins <i>in vitro</i> by 2DIR and TEM.

Abstract

(A, B) Overlay of diagonal slices for room temperature αB-crystallin (solid light blue) and heated sample (dashed light blue), or αB-R120G-crystallin (solid maroon) and heated sample (dashed maroon). The dotted vertical line indicates the amyloid-like β-sheet protein secondary structure peak frequency for αB-crystallin, as previously determined through TEM and 2DIR spectroscopy [10]. (C) Overlay of cross peaks for room temperature αB-crystallin (solid light blue) and heated sample (dashed light blue). (D) Overlay of cross peaks for room temperature αB-R120G-crystallin (solid maroon) and heated sample (dashed maroon). (E, F) TEM images for room temperature αB-crystallin (solid light blue) and heated sample (dashed light blue). White scale bars 100 nm. (G, H) TEM images for room temperature αB-R120G-crystallin (solid maroon) and heated sample (dashed maroon). White scale bar at 100 nm, dark blue scale bar at 200 nm. Room temperature samples were incubated at room temperature for 27 hours. Heated samples were heated for 2 hours at 43°C and then left at room temperature for 25 hours.</p