Figure 3: HTT subdomains bind HAP40 and can be used to reconstitute the HTT-HAP40 complex.
A, Size exclusion chromatography profiles of co-expressed NTD, CTD and HAP40 (black trace) after FLAG purification from Sf9 cells. An elution volume nearly identical to full-length HTT-HAP40 (orange trace) indicates formation of the trimeric complex. B, Reconstituted and full-length HTT-HAP40 complexes have similar melt profiles as determined by DSF. C, SDS-PAGE of full-length HTT (lane 1), NTD (lane 2), CTD (lane 3) and co-expressed subdomains (lane 4). D, Western blots of full-length HTT-HAP40, NTD, CTD, and reconstituted HTT-HAP40. E, Overlay, of cryo-EM map obtained from reconstituted HTT-HAP40 superimposed on the model generated for full-length HTT-HAP40 6X9O.
