Supplementary Table S1. Pathological analysis for non-synonymous sequence changes

Transcription

1 Abu-Amero and Bosley, Page 1 Supplementary Table S1. Pathological analysis for non-synonymous sequence changes 1393 G > A 3460 G > A 4040 C > G 4216 T > C This sequence change is located in the 12S rrna region and does not result in an amino acid change. It was found in 16.3% of Controls and was reported previously in normal individuals from multiple ethnic groups. Interspecies conservation was low. It was considered non-pathologic. This sequence change is located at codon 52 in the functional domain of the ND1 gene and changes a highly conserved Alanine, a hydrophobic amino acid (AA), to Threonine, a neutral AA, altering the hydropathy index from 0.66 to Protean predicted the introduction of a flexible region (flexibility score changed from 0 to 1.8). PolyPhen predicted this sequence change to be Probably Damaging, and SIFT predicted that it should affect protein function. This mutation is well established in the literature as a primary LHON mutation, 1-3 and it was not found in 159 Controls. This mutation is associated with LHON and was considered pathologic. This sequence change is located at amino acid position 245 in the transmembrane domain of the ND1 gene and changes a moderately conserved Threonine to a termination codon. Protean could not predict the consequence of this mutation since the change involves the introduction of a termination codon. PolyPhen predicted the change to be Probably Damaging, and SIFT predicted that it will affect protein function. However, a low heteroplasmy level of 18-20% (below the typical 60% threshold for pathologic mtdna mutations) and its presence in 6.3% of Controls raise doubts about its pathologic significance. It was considered non-pathologic. This sequence change is located at codon 304 in the transmembrane domain of the ND1 gene and changes a low conserved Tyrosine to a Histidine. Protean predicted no changes in hydropathy index or secondary structure. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it should not affect protein function. Although it has been reported as a secondary LHON mutation, it has been found in normal individuals 4 and was present in 46.5% of Controls. It was considered non-pathologic.

2 Abu-Amero and Bosley, Page A > G 4917 A > G 5319 A > T 6261 G > A This sequence change is located in pairing chain position 53 of the trna glutamine, just before the TψC loop. PolyPhen and SIFT databases could not predict the effect of this sequence change due to lack of data; however, phylogenetic data for 32 different species (Mamit-tRNA website; showed the sequence to be moderately conserved. This sequence alteration was not reported previously as a polymorphism and was absent in Controls. It was considered pathologic. This sequence change is located at codon 150 in the functional domain of the ND2 gene and changes a highly conserved Asparagine to Aspartic acid. Protean predicted no changes in hydropathy index or secondary structure. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it should not affect protein function. Although this sequence variant has been reported as a secondary LHON mutation, it has been detected in normal individuals 5, 6 and was present in 2.5% of Controls. It was considered non-pathologic. This sequence change is located at codon 284 in the functional domain of the ND2 gene and changes a moderately conserved Threonine to Serine. Protean predicted no changes in hydropathy index or secondary structure. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it should not affect protein function. It has been reported in an Indian lineage, 7 although it was not present in Controls here. This sequence variant was considered non-pathologic. This sequence variant is located at codon 120 in transmembrane domain of the cytochrome c oxidase subunit I (COI) which forms part of the catalytic subunit of the enzyme. It changes a moderately conserved Alanine (hydrophobic AA) to Threonine (neutral AA) and alters the hydropathy index from to Protean predicted the introduction of a flexible region as a result of this mutation. PolyPhen predicted this sequence change to be Possibly Damaging, and SIFT predicted that it should affect protein function. This mutation was previously detected in lymphocytes of six patients with prostate cancer, 8 where elevated levels of reactive oxygen species (ROS) implied possible pathogenicity. It was considered pathologic.

3 Abu-Amero and Bosley, Page C > G 7623 C > T 8656 A > T 8701 A > G This sequence variant is located at codon 489 in cytochrome c oxidase subunit I (COI). It changes of a low conserved Serine (neutral AA) to Cysteine (neutral AA), which is not expected to affect hydrophobicity of the protein. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it should not affect protein function. This sequence change has not been reported elsewhere as a polymorphism, and it was detected in 6.3% of Controls here. Heteroplasmy level was low (25%). It was considered non-pathologic. This sequence variant is located at codon 13 in cytochrome c oxidase subunit II (COII). It changes a highly conserved Threonine (neutral AA) to Isoleucine (Hydrophobic AA), which changed the hydropathy index from 0.21 to Protean predicted the loss of a flexible region as a result of this mutation. PolyPhen predicted this sequence change to be Possibly Damaging, and SIFT predicted that it should affect protein function. It was not reported previously as a polymorphism and was absent in Controls. It was considered pathologic. This sequence variant is located at codon 44 in the membrane proton channel CF(0) subunit of the ATP synthase 6 gene. It is located outside the conserved domain (extending from AA ) and changes a low conserved Threonine (neutral AA) to Serine (neutral AA). Protean predict no change in hydropathy index or secondary structure of the protein. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it should not affect protein function. This sequence change was not reported previously as a polymorphism and was absent in Controls. Nevertheless, it was considered non-pathologic. This sequence variant is located at amino acid position 59 outside the functional domain of ATPase 6 gene. It changes a low conserved Threonine with another low conserved Alanine, which resulted in a hydropathy index change from -0.7 to 1.8. Protean predicted no change in the protein structure. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it will not affect protein function. This sequence variant it is present in a number mitochondrial haplogroups and was found in 31.4% of Controls. It was considered non-pathologic.

4 Abu-Amero and Bosley, Page A > G 8860 A > G 9053 G > A This sequence variant is located at amino acid position 104 in the transmembrane conserved domain of the membrane proton channel CF(0) subunit of the ATP synthase A chain of ATPase 6. It changes a highly conserved Methionine to Valine, which may affect the translocation of protons across the membrane. Protean predicted substantial change in the protein structure with the disappearance of two Alpha regions and introduction of two new Beta regions. The hydropathy index changed from 1.60 to PolyPhen predicted this sequence change to be Probably Damaging, and SIFT predicted that it should affect protein function. This sequence alteration was reported previously but was absent in Controls here. It was considered pathologic. This sequence variant is located at amino acid position 112 in fragment 3 of the transmemebrane domain of the ATPase 6 gene. It changes a low conserved Threonine with another low conserved Alanine, which resulted in a hydropathy index change from -0.7 to 1.8. Protean predicted no change in the protein structure. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it will not affect protein function. This is a common sequence variant in a number of mitochondrial haplogroups, and it was found in 83.6% of Controls. It was considered non-pathologic. This sequence variant is located at amino acid position 176 outside the transmemebrane domain of the ATPase 6 gene. It changes a low conserved Serine with another low conserved Asparagine, which resulted in a hydropathy index change from -0.8 to Protean predicted no change in the protein structure as a result of this sequence change. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it will not affect protein function. This sequence variant is found in a number of mitochondrial haplogroups and was present in 31.4% of Controls. It was considered non-pathologic.

5 Abu-Amero and Bosley, Page G > A 9660 A > C 9667 A > G A > G This sequence variant is located at codon 177 in the conserved domain of the membrane proton channel CF(0) subunit of the ATP synthase A chain of ATPase 6. It changes a low conserved Alanine to Threonine. Protean predict no change in hydropathy index or secondary structure of the protein. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it should not affect protein function. This sequence variant was reported previously as a polymorphism 9 but was not detected in Controls here. It was considered non-pathologic. This sequence variant is located at amino acid position 152 in the functional domain (extending from AA 6 241) of cytochrome c oxidase subunit III (COIII). It changes a highly conserved Methionine to Leucine and may affect transfer of electrons from reduced cytochrome c to molecular oxygen. The hydropathy index changed from 1.83 to Protean predicted the loss of one Alpha region as a result of this mutation. PolyPhen predicted it to be Possibly Damaging, and SIFT predicted that it should affect protein function. This sequence change was not reported previously as a polymorphism and was absent in Controls. It was considered pathologic. This sequence variant is located at codon 154 in the functional domain of COIII and changes a lowly conserved Asparagine (Neutral) to Serine (Neutral). Protean did not predict a change in protein structure as a result of this sequence change. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it should not affect protein function. This sequence variant was detected previously in normal individuals 9 and in 1.9% of Controls here. It was considered non-pathologic. This sequence variant is located at amino acid position 114 outside the transmemebrane domain of the ND3 gene. It changes a low conserved Threonine with another low conserved Alanine, which resulted in a hydropathy index change from -0.7 to 1.8. Protean predicted no change in the protein structure as a result of this sequence change. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it will not affect protein function. This sequence variant is present in a number of mitochondrial haplogroups and was found in 66.7% of Controls. It was considered non-pathologic.

6 Abu-Amero and Bosley, Page A > G T > G T > C This sequence variant is located at amino acid position 25 of the catalytic domain (extending from AA 2 98) of NADH dehydrogenase 4L (ND4L). It changes a highly conserved Histidine to Arginine and may affect catalytic activity of the enzyme. This mutation caused the hydropathy index to change from 0.17 to 0.3. Protean predicted significant changes to the protein structure (loss of two Alpha regions and introduction of new Beta regions and a flexible region). PolyPhen predicted this sequence change to be Probably Damaging, and SIFT predicted that it should affect protein function. This sequence alteration was not reported previously as a polymorphism, was absent in Controls here, and was heteroplasmic at a relatively high level (70%). It was considered pathologic. This sequence variant is located at amino acid position 41 of the catalytic transmembrane domain of ND4L. It changes a highly conserved Phenylalanine to Cysteine and may affect catalytic activity of the enzyme. The hydropathy index changed from 2.11 to Protean predicted introduction of a new Beta region. PolyPhen predicted this sequence change to be Probably Damaging, and SIFT predicted that it should affect protein function. This sequence was reported previously as a somatic mutation, 10 was absent in Controls here, and was heteroplasmic at a relatively high level (75%). It was considered pathologic. This sequence change is located at codon 65 in the transmembrane conserved domain of ND4L and changes a highly conserved Valine to Alanine (neutral AA). Hydropathy index changed from to Protean predicted no structural changes as a result of this mutation. PolyPhen predicted this sequence change to be Probably Damaging, and SIFT predicted that it should affect protein function. This nucleotide change was reported previously as a provisional LHON mutation 11 and was absent in Controls here. It was considered pathologic.

7 Abu-Amero and Bosley, Page G > A C > A C > T This sequence change is located at codon 340 in the functional domain (extending from AA ) of the ND4 gene and changes a highly conserved Arginine to Histidine. The hydropathy index changed from 0.11 to Protean predicted no change in the protein structure as a result of this mutation. PolyPhen predicted this sequence change to be Probably Damaging, and SIFT predicted that it will affect protein function. It is well established as a primary LHON mutation responsible for the majority of LHON cases reported worldwide and was considered pathologic. This sequence variant is located at amino acid position 372 in the transmembrane functional conserved domain of ND4 and changes a highly conserved Threonine to Asparagine. This protein is part of complex I, which catalyzes the transfer of two electrons from NADH to ubiquinone, and a pathologic mutation in this location may affect catalytic activity. The hydropathy index changed from 1.14 to Protean predicted no change to the protein structure. PolyPhen predicted this sequence change to be Possibly Damaging, and SIFT predicted that it should affect protein function. This sequence alteration was not reported previously as a polymorphism and was absent in Controls here. It was considered pathologic. This sequence variant is located at amino acid position 4 outside the transmemebrane domain of the ND5 gene and changes a low conserved Histidine with another low conserved Tyrosine. Protean predicted no change in the protein structure as a result of this sequence change. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it will not affect protein function. This sequence variant is found in a number of mitochondrial haplogroups and was present in 32.7% of Controls. It was considered non-pathologic.

8 Abu-Amero and Bosley, Page C > T T > G A > G This sequence variant is located at amino acid position 23 outside the transmembrane domain of the ND5 gene. It changes a low conserved Leucine with another low conserved Phenylalanine, which resulted in a hydropathy index change from 3.8 to 2.8. Protean predicted no change in the protein structure because of this sequence change. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it will not affect protein function. This sequence variant is present in a number of mitochondrial haplogroups and was found in 35.2% of Controls. It was considered non-pathologic. This sequence variant is located at amino acid position 149 in the transmembrane conserved domain (oxidoreductase domain) of NADH dehydrogenase subunit 5 (ND5). It changes a highly conserved Isoleucine (hydrophobic AA) to Serine (Neutral AA), which changed the hydropathy index from 1.34 to Protean predicted the disappearance of two Beta regions and the introduction of a new turn and a new coil region, which may affect the catalytic activity of the enzyme. PolyPhen predicted this sequence change to be Probably Damaging, and SIFT predicted that it should affect protein function. This sequence alteration was not reported previously as a polymorphism, was absent in Controls here, and was heteroplasmic at a relatively high level (65%). It was considered pathologic. This sequence variant is located at amino acid position 205 outside the transmembrane domain of the ND5 gene. It changes a low conserved Asparagine with another low conserved Serine, which resulted in a hydropathy index change from -3.5 to 0.8. Protean predicted no change in the protein structure because of this sequence change. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it will not affect protein function. This sequence variant is present in a number of mitochondrial haplogroups and was found in 41.5% of Controls. It was considered non-pathologic.

9 Abu-Amero and Bosley, Page A > C A > G G > A This sequence variant is located at amino acid position 348 in the conserved domain of ND5, where a pathologic mutation might affect catalytic activity of the enzyme. It changes a moderately conserved Histidine (hydrophilic AA) to Proline (hydrophobic AA), which changed the hydropathy index from 0.23 to Protean predicted the loss of a Beta region and the gain of a new turn. PolyPhen predicted this sequence change to be Probably Damaging, and SIFT predicted that it should affect protein function. It was not reported previously as a polymorphism and was absent in Controls here. It was considered pathologic. This sequence variant is located at amino acid position 449 of ND5, outside the functional domain and near the C terminus, where it changes a lowly conserved Threonine to Alanine. Protean predicted no change to the protein structure. The hydropathy index changed from 1.14 to PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it should not affect protein function. This sequence change was reported previously as a polymorphism but was absent in Controls here. It was considered non-pathologic. This sequence variant is located at codon 458 in the ND5 C terminus and changes a moderately conserved Alanine (Hydrophobic) to Threonine (Neutral), which changes the hydropathy index from 0.68 to Protean predicted the introduction of a new flexible region as a result of this mutation. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it should not affect the protein function. This sequence variant was reported previously in LHON patients as well as healthy individuals 5 and was detected in 46.5% of Controls here. It was considered nonpathologic.

10 Abu-Amero and Bosley, Page C > T T > C A > G T > C This sequence variant is located at amino acid position 533 outside the transmembrane domain of the ND5 gene. It changes a low conserved Threonine with another low conserved Methionine, which resulted in a hydropathy index change from -0.7 to 1.9. Protean predicted no change in the protein structure because of this sequence change. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it will not affect protein function. This sequence variant is present in some mitochondrial haplogroups and was found in 28.9% Controls. It was considered non-pathologic. This sequence variant is located at amino acid position 592 in the transmembrane domain of the ND5 gene. It changes a low conserved Phenylalanine with another low conserved Leucine, which resulted in a hydropathy index change from 2.8 to 3.8. Protean predicted no change in the protein structure because of this sequence change. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it will not affect protein function. This sequence variant is present in some mitochondrial haplogroups and was found in 16.3% of Controls. It was considered non-pathologic. This sequence variant is located at amino acid position 31 in the transmembrane segment of the NADH dehydrogenase subunit 6 (ND6) and changes a lowly conserved Valine to Alanine. The hydropathy index changed from 1.96 to 1.69, and Protean predicted no change to the protein structure. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it should not affect protein function. This sequence change was reported previously as a polymorphism and was found in 2.5% of Controls here. It was considered non-pathologic. This sequence variant is located at amino acid position 7 in the N-terminal domain of the Cytochrome B gene. It changes a low conserved Isoleucine with another low conserved Threonine. Protean predicted no change in the protein structure because of this sequence change. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it will not affect protein function. This sequence change was reported previously as a polymorphism and was found in 74.8% Controls. It was considered non-pathologic.

11 Abu-Amero and Bosley, Page T > C C > T G > A T > C This sequence variant is located at codon 18 in the N-terminal domain of Cytochrome b and changes a highly conserved Phenylalanine to Leucine. The hydropathy index changed from 0.33 to 0.44, and Protean predicted no change in the protein structure. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it should not affect protein function. This sequence change was reported previously as a polymorphism and was found in 1.2% of Controls. It was considered non-pathologic. This sequence variant is located at codon 39 in the N-terminal domain of Cytochrome b (CYTB) and changes a low conserved Alanine to Valine. Hydropathy index changed from 2.53 to 0.28, and Protean predicted the introduction of a new Beta region. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it should not have an affect on protein function. This sequence change was reported previously as a polymorphism and was found in 32.7% of Controls here. It was considered non-pathologic. This sequence variant is located at codon 171 in the N-terminal domain of CYTB and changes a highly conserved Aspartic acid to Asparagine. It causes no change in the hydropathy index (0.12), and Protean predicted no change in the protein structure. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it should not have an affect on protein function. This sequence variant has been reported in certain LHON patients 17, 18 and in healthy individuals, 19 and it was present in 7.5% of Controls. It was considered non-pathologic. This sequence variant is located at amino acid position 194 in the N-terminal domain of Cytochrome B gene. It changes a low conserved Threonine with another low conserved Alanine. Protean predicted no change in the protein structure because of this sequence change. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it will not affect protein function. This sequence variant is present in a number of mitochondrial haplogroups and was found in 72.9% of Controls. It was considered non-pathologic.

12 Abu-Amero and Bosley, Page C > A T > C This sequence variant is located at amino acid position 236 in the transmembrane domain of cytochrome B gene and changes a low conserved Leucine with another low conserved Isoleucine. Protean predicted no change in the protein structure because of this sequence change. PolyPhen predicted this sequence change to be Benign, and SIFT predicted that it will not affect protein function. This sequence variant is present in a number of mitochondrial haplogroups and was found in 61% of Controls. It was considered non-pathologic. This sequence variant is located at amino acid position 310 in the C-terminal domain of CYTB, where a pathologic mutation may affect ubiquinol/ubiquinone binding activity. It changes a highly conserved Serine (neutral AA) to Proline (hydrophobic AA), which altered the hydropathy index from 0.61 to Protean predicted no change to the protein structure. PolyPhen predicted this sequence change to be Possibly Damaging, and SIFT predicted that it should affect protein function. This sequence alteration has not been reported previously as a polymorphism and was absent in Controls here. It was considered pathologic. Pathogenicity was assessed using standard criteria 20 and previous reports of sequence variants found in the MITOMAP database, the Human Mitochondrial Genome Database ( GenBank ( and Medline listed publications. Interspecies conservation was assessed using the PolyPhen (Polymorphism Phenotyping) database ( which determines interspecies conservation for an altered amino acid by performing alignments with all available amino acid sequences for other species, and, when necessary, using the Mamit-tRNA website ( PolyPhen pathogenicity prediction was assessed utilizing the PolyPhen database. Probably Damaging constitutes a high confidence of affecting protein function or structure. Possibly Damaging reflects a likelihood of affecting protein function or structure, while Benign changes most likely lack phenotypic effect. Unknown means that PolyPhen could make no prediction due to lack of data. SIFT (Sorting Intolerant from Tolerant; returns predictions for which amino acid

13 Abu-Amero and Bosley, Page 13 s will affect protein function given a particular protein sequence. Protean (Protein Structure Prediction and Annotation) is part of the LASERGENE V.6 software (DNASTAR, Inc. Madison, WI, USA). Protean helps to predict and display patterns, secondary structural characteristics, and physiochemical properties (hydropathy index, flexibility index and antigenic index). The Hydropathy Index was measured by Protean according to the Kyte-Doolittle method, 21 which predicts the regional hydropathy of proteins from their amino acid sequence. Hydropathy values were assigned for all amino acids and were then averaged over a window size = 7. Results below 0 are hydrophobic and above 0 are hydrophilic. Flexibility Index was measured by Protean according to the method of Karplus and Schultz, 22 which predict backbone chain flexibility. Residues with values below 1 are rigid and above 1 are flexible. The most flexible residues are assigned a value of 2 and the least flexible is assigned 0. Pathogenic mutations commonly involve the trans-membrane domains. 23 References 1.Bose S, Dhillon N, Ross-Cisneros FN, Carelli V. Relative post-mortem sparing of afferent pupil fibers in a patient with 3460 Leber's hereditary optic neuropathy. Graefes Arch Clin Exp Ophthalmol 2005: Cock HR, Cooper JM, Schapira AH. Functional consequences of the 3460-bp mitochondrial DNA mutation associated with Leber's hereditary optic neuropathy. J Neurol Sci 1999;165(1): Hwang JM, Chang BL, Koh HJ, et al. Leber's hereditary optic neuropathy with 3460 mitochondrial DNA mutation. J Korean Med Sci 2002;17(2): Chinnery PF, Turnbull DM, Howell N, Andrews RM. Mitochondrial DNA mutations and pathogenicity. J Med Genet 1998;35(8): Brown MD, Torroni A, Reckord CL, Wallace DC. Phylogenetic analysis of Leber's hereditary optic neuropathy mitochondrial DNA's indicates multiple independent occurrences of the common mutations. Hum Mutat 1995;6(4): Howell N, Kubacka I, Halvorson S, et al. Phylogenetic analysis of the mitochondrial genomes from Leber hereditary optic neuropathy pedigrees. Genetics 1995;140(1):

14 Abu-Amero and Bosley, Page 14 7.Rajkumar R, Banerjee J, Gunturi HB, et al. Phylogeny and antiquity of M macrohaplogroup inferred from complete mt DNA sequence of Indian specific lineages. BMC Evol Biol 2005;5(1):26. 8.Petros JA, Baumann AK, Ruiz-Pesini E, et al. mtdna mutations increase tumorigenicity in prostate cancer. Proc Natl Acad Sci U S A 2005;102(3): Finnila S, Hassinen IE, Majamaa K. Phylogenetic analysis of mitochondrial DNA in patients with an occipital stroke. Evaluation of mutations by using sequence data on the entire coding region. Mutat Res 2001;458(1-2): Taylor RW, Barron MJ, Borthwick GM, et al. Mitochondrial DNA mutations in human colonic crypt stem cells. J Clin Invest 2003;112(9): Brown MD, Starikovskaya E, Derbeneva O, et al. The role of mtdna background in disease expression: a new primary LHON mutation associated with Western Eurasian haplogroup J. Hum Genet 2002;110(2): Rufa A, Dotti MT, Cardaioli E, et al. Leber hereditary optic neuropathy in 2 of 4 siblings with mtdna mutation: clinical variability or effect of toxic environmental exposure? Eur Neurol 2005;53(1): Mashima Y, Hiida Y, Oguchi Y, et al. High frequency of mutations at position in mitochondrial ND4 gene in Japanese families with Leber's hereditary optic neuropathy. Hum Genet 1993;92(1): Barboni P, Mantovani V, Montagna P, et al. Mitochondrial DNA analysis in Leber's hereditary optic neuropathy. Ophthalmic Paediatr Genet 1992;13(4): Kormann BA, Schuster H, Berninger TA, Leo-Kottler B. Detection of the G to A mitochondrial DNA mutation at position in German families with Leber's hereditary optic neuropathy. Hum Genet 1991;88(1): Cortelli P, Montagna P, Avoni P, et al. Leber's hereditary optic neuropathy: genetic, biochemical, and phosphorus magnetic resonance spectroscopy study in an Italian family. Neurology 1991;41(8):

15 Abu-Amero and Bosley, Page Johns DR, Neufeld MJ. Pitfalls in the molecular genetic diagnosis of Leber hereditary optic neuropathy (LHON). Am J Hum Genet 1993;53(4): Johns DR, Smith KH, Savino PJ, Miller NR. Leber's hereditary optic neuropathy. Clinical manifestations of the mutation. Ophthalmology 1993;100(7): Hofmann S, Bezold R, Jaksch M, et al. Disease relevance of the so-called secondary Leber hereditary optic neuropathy mutations. Am J Hum Genet 1997;60(6): Chinnery PF, Howell N, Andrews RM, Turnbull DM. Mitochondrial DNA analysis: polymorphisms and pathogenicity. J Med Genet 1999;36(7): Kyte J, Doolittle RF. A simple method for displaying the hydropathic character of a protein. J Mol Biol 1982;157(1): Karplus PA, Schultz GE. Prediction of chain flexibility in proteins. Naturwissenschaft 1985;72: Mitchell AL, Elson JL, Howell N, et al. Sequence Variation In Mitochondrial Complex I Genes: Mutation Or Polymorphism? J Med Genet 2005.