title: Misfolding, degradation, and aggregation of variant proteins. The molecular pathogenesis of short chain acyl-CoA dehydrogenase (SCAD) deficiency.
abstract: Short chain acyl-CoA dehydrogenase (SCAD) deficiency is an inborn error of the mitochondrial fatty acid metabolism caused by rare variations as well as common susceptibility variations in the SCAD gene. Earlier studies have shown that a common variant SCAD protein (R147W) was impaired in folding, and preliminary experiments suggested that the variant protein displayed prolonged association with chaperonins and delayed formation of active enzyme. Accordingly, the molecular pathogenesis of SCAD deficiency may rely on intramitochondrial protein quality control mechanisms, including degradation and aggregation of variant SCAD proteins. In this study we investigated the processing of a set of disease-causing variant SCAD proteins (R22W, G68C, W153R, R359C, and Q341H) and two common variant proteins (R147W and G185S) that lead to reduced SCAD activity. All SCAD proteins, including the wild type, associate with mitochondrial hsp60 chaperonins; however, the variant SCAD proteins remained associated with hsp60 for prolonged periods of time. Biogenesis experiments at two temperatures revealed that some of the variant proteins (R22W, G68C, W153R, and R359C) caused severe misfolding, whereas others (R147W, G185S, and Q341H) exhibited a less severe temperature-sensitive folding defect. Based on the magnitude of in vitro defects, these SCAD proteins are characterized as folding-defective variants and mild folding variants, respectively. Pulse-chase experiments demonstrated that the variant SCAD proteins either triggered proteolytic degradation by mitochondrial proteases or, especially at elevated temperature, aggregation of non-native conformers. The latter finding may indicate that accumulation of aggregated SCAD proteins may play a role in the pathogenesis of SCAD deficiency.
mesh_terms: D000818:Animals; D001081:Apyrase / Q000737:chemistry; D015153:Blotting, Western; D044926:Butyryl-CoA Dehydrogenase / Q000737:chemistry* / Q000172:deficiency* / Q000378:metabolism; D018834:Chaperonin 60 / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D014644:Genetic Variation; D005838:Genotype; D008052:Lipid Metabolism, Inborn Errors / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D008928:Mitochondria / Q000378:metabolism; D008954:Models, Biological; D008958:Models, Molecular; D017830:Octoxynol / Q000494:pharmacology; D009840:Oligomycins / Q000494:pharmacology; D010957:Plasmids / Q000378:metabolism; D014176:Protein Biosynthesis; D011487:Protein Conformation; D017510:Protein Folding; D013696:Temperature; D013997:Time Factors; D014158:Transcription, Genetic
chemical_list: D018834:Chaperonin 60; D018076:DNA, Complementary; D003902:Detergents; D009840:Oligomycins; D017830:Octoxynol; D044926:Butyryl-CoA Dehydrogenase; D001081:Apyrase