Fig. 3. The R207C-I138C double-cysteine PrP^C mutant acts as an H-latch mimic.

a,b, ¹⁵N-heteronuclear single quantum coherence spectra of rmPrP free (red) and mPrP^2cys (blue). Residues with different chemical shifts in the two spectra are colored orange on the GD structure in b, which resemble the H-latch conformation in the POM1–PrP complex. c, MD simulations show that mPrP^2cys resembles the PrP–POM1 complex, with increased flexibility in the α2–α3 and β2–α2 loops and decreased flexibility in the 2Cys region, corresponding to the POM1 epitope. d–f, Prnp^ZH3/ZH3 COCS transduced with a bi-cistronic AAV expressing mNG and mPrP^C (left) or mPrP^2Cys (right). See Extended Data Figure 3f for quantification. Scale bars: 250 µm. d, mNG was visible in all COCS at 15 days post transduction (dpt, top row) but disappeared in mPrP^2Cys at 31 dpt (bottom row). e, Calbindin-1⁺ Purkinje cells were preserved at 15 dpt but became largely undetectable at 31 dpt, possibly as a result of mPrP^2Cys toxicity. f, Dose escalation of twice as many viral vectors as in d and e led to earlier onset of mPrP^2cys-mediated neurodegeneration. Significant neurodegeneration was observable at 15 dpt; see quantification in Extended Data Figure 3f.