Secondary structure analysis of MAK33 V_L in the fibril state.

<p>A) β-sheet propensity calculated with TALOS+ [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0181799#pone.0181799.ref032" target="_blank">32</a>]. B) Sequence and secondary structure elements of the native V_L fold. Green and red bars indicate β-strands and CDRs of the native structure, respectively. Red arrows below the sequence indicate β-strands in the fibril state. The expansion shows the assigned atoms in the aggregated state.</p

Comparison of MAK33 V_L oligomers and fibrils.

<p>A) Procedure to form oligomers and fibrils. B), C) Electron micrographs of MAK33 V_L S20N oligomers (B) and fibrils (C). The scale bar denotes 200 nm. D) FTIR spectra of MAK33 V_L S20N oligomers and fibrils. The peak maxima were 1619 cm^-1 (oligomers) and 1621 cm^-1 (fibrils), respectively. The oligomer spectrum displayed an additional peak at 1697 cm^-1. E) PDSD ¹³C,¹³C-intraresidue correlations of MAK33 V_L S20N fibrils and MAK33 V_L WT oligomers. The proline spin system, which is more intense in the oligomers, is highlighted in blue.</p

¹³C,¹⁵N correlations of MAK33 V_L S20N fibrils.

<p>A) N(CA)CX spectrum of a u-¹³C,¹⁵N labeled sample. B) NCA spectrum of a 2-¹³C-glycerole isotope labeled sample. Peak positions are identical in A) and B), indicating good reproducibility. The resolution in B) is increased due to sparse isotope labeling.</p

Comparison with AL-09, amyloid prediction algorithms and native state chemical shifts.

<p>A) Sequence alignment of MAK33 V_L S20N and AL-09 V_L: Identical residues are marked in blue. Residues assigned in MAS ssNMR spectra are indicated by bars above and below the corresponding sequence. B) Predictions of MAK33 V_L S20N amyloid propensity and experimentally observed β-strands. C) Secondary chemical shift correlation of MAK33 V_L S20N in the solid-state (fibrils, pH 2) and solution-state (native, pH 6.5) for Cα, Cβ, CO and N chemical shifts. The cross-correlation coefficients r are indicated in each plot.</p