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. 2011 Feb 7;192(3):447–462. doi: 10.1083/jcb.201008177

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© 2011 Claypool et al.

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Figure 6. — i-AAA protease mutants bind the A88E BTHS mutant, but not wt, tafazzin. (A) Steady-state expression was determined from whole-cell extracts derived from Δtaz1Δyme1 yeast transformed with the indicated Taz1p and Yme1p variants by immunoblotting for Taz1p, Yme1p, and the loading control, porin. (B) Digitonin extracts from mitochondria derived from the indicated strains were subjected to Ni²⁺ nitrilotriacetic acid (NiNTA) chromatography. The indicated amount of total, TCA-precipitated flow through, and bound material was resolved by SDS-PAGE and immunoblotted as indicated. The asterisk highlights the cross-reaction with porin of the AAC2 antiserum. (C) Digitonin extracts from mitochondria derived from the indicated strains were resolved by 2D BN/SDS-PAGE, and Taz1p complexes were detected by immunoblotting. 150 µg Δtaz1Δyme1 yeast transformed with empty vector or Yme1 mutants and 250 µg Δtaz1Δyme1 (Wt Yme1) were analyzed. The four BTHS mutants being characterized in the present study are shown in red. Stds, molecular weight standards. (A–C) n = 3.