TY - JOUR
T1 - Mutational hotspots in electron transfer flavoprotein underlie defective folding and function in multiple acyl-CoA dehydrogenase deficiency
AU - Gomes, Claudio Emanuel
N1 - Henriques, B
PY - 2010/1/1
Y1 - 2010/1/1
N2 - We have carried out an extensive in silico analysis on 18 disease associated missense mutations found in electron transfer flavoprotein (ETF), and found that mutations fall essentially in two groups, one in which mutations affect protein folding and assembly, and another one in which mutations impair catalytic activity and disrupt interactions with partner dehydrogenases. We have further experimentally analyzed three of these mutations, ETF beta-p Cys42Arg, ETF beta-p.Asp128Asn and ETF beta-p.Arg191Cys, which have been found in homozygous form in patients and which typify different scenarios in respect to the clinical phenotypes The ETF beta-p Cys42Arg mutation, related to a severe form of multiple acyl-CoA dehydrogenase deficiency (MADD), affects directly the AMP binding site and intersubunit contacts and impairs correct protein folding. The two other variations, ETF beta-p Asp128Asn and ETF beta-p.Arg191Cys, are both associated with mild MADD, but these mutations have a different impact on ETF. Although none affects the overall alpha/beta fold topology as shown by far-UV CD, analysis of the purified proteins shows that both have substantially decreased enzymatic activity and conformational stability. Altogether, this study combines in silica analysis of mutations with experimental data and has allowed establishing structural hotspots within the ETF fold that are useful to provide a rationale for the prediction of effects of mutations in ETF.
AB - We have carried out an extensive in silico analysis on 18 disease associated missense mutations found in electron transfer flavoprotein (ETF), and found that mutations fall essentially in two groups, one in which mutations affect protein folding and assembly, and another one in which mutations impair catalytic activity and disrupt interactions with partner dehydrogenases. We have further experimentally analyzed three of these mutations, ETF beta-p Cys42Arg, ETF beta-p.Asp128Asn and ETF beta-p.Arg191Cys, which have been found in homozygous form in patients and which typify different scenarios in respect to the clinical phenotypes The ETF beta-p Cys42Arg mutation, related to a severe form of multiple acyl-CoA dehydrogenase deficiency (MADD), affects directly the AMP binding site and intersubunit contacts and impairs correct protein folding. The two other variations, ETF beta-p Asp128Asn and ETF beta-p.Arg191Cys, are both associated with mild MADD, but these mutations have a different impact on ETF. Although none affects the overall alpha/beta fold topology as shown by far-UV CD, analysis of the purified proteins shows that both have substantially decreased enzymatic activity and conformational stability. Altogether, this study combines in silica analysis of mutations with experimental data and has allowed establishing structural hotspots within the ETF fold that are useful to provide a rationale for the prediction of effects of mutations in ETF.
KW - GENOTYPE
KW - FRATAXIN
KW - DISEASES
KW - ACIDEMIA TYPE-II
KW - PHENOTYPE
KW - STABILITY
KW - VARIANT
KW - OXIDATION
KW - ALPHA-SUBUNIT
KW - RIBOFLAVIN
U2 - 10.1016/j.bbadis.2010.07.015
DO - 10.1016/j.bbadis.2010.07.015
M3 - Article
VL - 1802
SP - 1070
EP - 1077
JO - Biochimica Et Biophysica Acta-Molecular Basis Of Disease
JF - Biochimica Et Biophysica Acta-Molecular Basis Of Disease
SN - 0925-4439
IS - 11
ER -