Figure 1.

Mzt1 Prevents Large-Scale Aggregation of the γ-TuSC In Vitro

(A) SDS-PAGE of γ-TuSC^Alp6-MBP (without Mzt1), purified on amylose via Alp6-MBP.

(B) Superose 6 size-exclusion chromatography (SEC) of γ-TuSC^Alp6-MBP, with corresponding SDS-PAGE of indicated fractions. The majority of eluting γ-TuSC^Alp6-MBP elutes in the void volume.

(C) SDS-PAGE of 80-min density-gradient centrifugation of γ-TuSC^Alp6-MBP (SYPRO Ruby stain). Most γ-TuSC is in the pellet. Results from a 45-min centrifugation are shown in Figure S1C.

(D) SDS-PAGE of γ-TuSC^Alp6-MBP with Mzt1 (γ-TuSC^Alp6-MBP:Mzt1), purified as above. Mzt1 is shown from a higher-contrast image, shown in Figure S1B.

(E) Superose 6 SEC of (γ-TuSC^Alp6-MBP:Mzt1), with corresponding SDS-PAGE of indicated fractions. Alp6-MBP, Alp4, γ-tubulin (Gtb1), and Mzt1 coelute, much later than the void volume.

(F) SDS-PAGE of 80-min density-gradient centrifugation of γ-TuSC^Alp6-MBP:Mzt1 (SYPRO Ruby stain). γ-TuSC^Alp6-MBP:Mzt1 sediments with a broad profile centered at ∼24S. Mzt1 was visualized on a separate gel with higher acrylamide concentration. Results from a 45-min centrifugation are shown in Figure S1D.

See also Figure S1.