A molecular chaperone for mitochondrial complex I assembly is mutated in a progressive encephalopathy
J. Clin. Invest. Isla Ogilvie, et al. 115:2784 doi:10.1172/JCI26020 [
Go to this article.]
Figure 5Analysis of expression of B17.2L by BN-PAGE immunoblot. (
A and
B) Mitochondria from control (lane C) and patient (lanes Be, Br, and A) muscle were subjected to first-dimension BN-PAGE analysis. The gel was immunoblotted and reacted with antibodies directed against ND1 (
A) or B17.2L (
B). The lower panel in
A is an immunoblot of the loading control (70-kDa subunit of complex II). (
C) BN-PAGE analysis of complex I assembly in patient fibroblasts. Mitoplasts were isolated from control and patient fibroblasts and subjected to second-dimension BN-PAGE analysis. A cocktail of antibodies directed against the indicated proteins was used to probe the second-dimension immunoblots. The lines labeled 1–3 on the gel indicate the position of the complex I-III supercomplex, fully assembled complex I, and the 830-kDa–complex III subcomplex, respectively. The spot that runs above the 49-kDa subunit and intersects with line 3 is a nonspecific protein recognized by the anti–49-kDa antibody. BI7.2L runs at its approximate monomeric MW in control and patient F (smear at the extreme right of the gel) and with an 830-kDa subcomplex of complex I in the patients with the known structural subunit gene mutations (Be and Bo). (
D) First-dimension BN-PAGE gel demonstrating the complete absence of fully assembled complex I in muscle mitochondria from patient Bo.
