Selective association of misfolded ALS-linked mutant SOD1 with the cytoplasmic face of mitochondria

*Ludwig Institute and Departments of Medicine and Neuroscience, University of California at San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0670; and
^§Department of Medicine (Neurology) and Brain Research Centre, University of British Columbia Hospital, University of British Columbia, 2211 Wesbrook Mall, Vancouver, BC, Canada V6T 2B5

Contributed by Don W. Cleveland, December 27, 2007 (received for review December 10, 2007)

Abstract

Mutations in copper/zinc superoxide dismutase (SOD1) are causative for dominantly inherited amyotrophic lateral sclerosis (ALS). Despite high variability in biochemical properties among the disease-causing mutants, a proportion of both dismutase-active and -inactive mutants are stably bound to spinal cord mitochondria. This mitochondrial proportion floats with mitochondria rather than sedimenting to the much higher density of protein, thus eliminating coincidental cosedimentation of protein aggregates with mitochondria. Half of dismutase-active and ≈90% of dismutase-inactive mutant SOD1 is bound to mitochondrial membranes in an alkali- and salt-resistant manner. Sensitivity to proteolysis and immunoprecipitation with an antibody specific for misfolded SOD1 demonstrate that in all mutant SOD1 models, misfolded SOD1 is deposited onto the cytoplasmic face of the outer mitochondrial membrane, increasing antigenic accessibility of the normally structured electrostatic loop. Misfolded mutant SOD1 binding is both restricted to spinal cord and selective for mitochondrial membranes, implicating exposure to mitochondria of a misfolded mutant SOD1 conformer mediated by a unique, tissue-selective composition of cytoplasmic chaperones, components unique to the cytoplasmic face of spinal mitochondria to which misfolded SOD1 binds, or misfolded SOD1 conformers unique to spinal cord that have a selective affinity for mitochondrial membranes.

Footnotes

^¶To whom correspondence should be addressed. E-mail: dcleveland{at}ucsd.edu
Author contributions: C.V.V. and D.W.C. designed research; C.V.V. and T.M.M. performed research; N.R.C. contributed new reagents/analytic tools; C.V.V. and D.W.C. analyzed data; and C.V.V. and D.W.C. wrote the paper.
↵ ^†Present address: Department of Medicine, CHUM Research Center, University of Montréal, 1560 rue Sherbrooke Est, Montréal, QC, Canada H2L 4M1.
↵ ^‡Present address: Department of Neurology, Hope Center for Neurological Disorders, Washington University, School of Medicine, 660 South Euclid Avenue, St. Louis, MO 63110.
The authors declare no conflict of interest.
This article contains supporting information online at www.pnas.org/cgi/content/full/0712209105/DC1.