Figure 3.
MPC Subunits Are Imported along the Carrier Pathway
(A) Isolated mitochondria from WT and tim22-14 cells grown for 15 h at the non-permissive temperature (37°C) were solubilized and analyzed by BN-PAGE and immunoblotting.
(B and C) [35S]-labeled Mpc1 was imported into (B) WT and mpc1Δ mitochondria and (C) mpc1Δ, tim22-14/mpc1Δ, and tim10-2/mpc1Δ mitochondria.
(D) [35S]-labeled Atp5 was imported into mpc1Δ and tim22-14/mpc1Δ mitochondria.
(E) Membrane potential measurement of mpc1Δ, tim22-14/mpc1Δ, and tim10-2/mpc1Δ mitochondria.
(F and G) [35S]-labeled Mpc3 was imported into (F) WT and tim22-14 mitochondria and (G) mpc1Δ and tim22-14/mpc1Δ mitochondria.
(H) [35S]-labeled Mpc2 was imported into WT and tim22-14 mitochondria.
(I) Purified mitochondria from WT and Tim22-downregulated (Tim22↓) cells were analyzed by SDS-PAGE and western blotting.
(J) Membrane potential measurement of WT and Tim22-downregulated mitochondria.
(K and L) [35S]-labeled (K) Mpc2 and (L) Crc1 were imported into WT and Tim22-downregulated mitochondria. In all import experiments, analyses were carried out by BN-PAGE and digital autoradiography. Δψ, membrane potential.
See also Figure S3.