Fig. 4: Dynamic nature of NET1ΔC - SEC-MALS and analytical ultracentrifugation. | Nature Communications

Fig. 4: Dynamic nature of NET1ΔC - SEC-MALS and analytical ultracentrifugation.

From: The dynamic nature of netrin-1 and the structural basis for glycosaminoglycan fragment-induced filament formation

Fig. 4

a SEC-MALS analysis of NET1ΔC in absence of HOs shows an equilibrium between monomeric and dimeric NET1ΔC. Upon the addition of HO-dp8, HO-dp10 and HO-dp12 we see a step-wise increase of mass with growing HO length. In the presence of HO-dp20, NET1ΔC forms a higher-order oligomer with a calculated mass of 346 kDa. Black lines show the mass distributions across the elution peaks which were traced by UV (A280 nm, cyan), differential refractive index (dRI658 nm, gold) and light scattering at 90° angle (665 nm, red). b c(s, M) distribution obtained from sedimentation velocity at 13 µM concentration without the addition of HOs reveal a monomer-dimer equilibrium (top). In the presence of HO-dp8 (1:1 and 1:2 molar ratios), NET1ΔC forms populations of particles with a stepwise increasing mass of ~160 kDa that form a ladder with an upper mass limit of 729 ± 49 kDa. Exact oligomer assignment is ambiguous for the larger species (Supplementary Tables 3 and 4).

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