cge12316-sup-0001-FigureS1.docxWord 2007 document17KFig. S1. Multiple sequence alignment (MSA) of human ASAH1 with homologues from other species. The MSA was performed as described in the 'Materials and methods' section. The amino acid position is shown at the beginning and end of the sequence; each mutated residue is indicated by an arrow. The homologues are as follows: gi|189011548|ref|NP_808592.2| Homo sapiens; gi|148727296|ref|NP_001092030.1| Pan troglodytes; gi|332215273|ref|XP_003256766.1| Nomascus leucogenys; gi|109085733|ref|XP_001098342.1| Macaca mulatta; gi|55730315|emb|CAH91880.1| Pongo abelii; gi|62510479|sp|Q60HH4.1| Macaca fascicularis; gi|390473621|ref|XP_002756937.2| Callithrix jacchus; gi|301776827|ref|XP_002923833.1| Ailuropoda melanoleuca; gi|350594565|ref|XP_003134235.3| Sus scrofa; gi|397506342|ref|XP_003823688.1| Pan paniscus; gi|186972122|ref|NP_001068927.1| Bos Taurus; gi|354470785|ref|XP_003497625.1| Cricetulus griseus; gi|40254747|ref|NP_445859.2| Rattus norvegicus; gi|9790019|ref|NP_062708.1| Mus musculus; gi|348561387|ref|XP_003466494.1| Cavia porcellus; gi|41055548|ref|NP_956871.1| Danio rerio; gi|57530079|ref|NP_001006453.1| Gallus gallus; gi|115712075|ref|XP_784832.2| Strongylocentrotus purpuratus; gi|268561274|ref|XP_002646404.1| Caenorhabditis briggsae; gi|17508209|ref|NP_493173.1| Caenorhabditis elegans; gi|308464848|ref|XP_003094688.1| Caenorhabditis remanii; gi|348524526|ref|XP_003449774.1| Oreochromis niloticus.
cge12316-sup-0002-FigureS2.docxWord 2007 document17KFig. S2. Alignment of templates 2X1D_A with the AC α-subunit (Met1 to Ile142) and of 2BJF_A and 3GVZ_A with the β-subunit (Cys143 to Trp395) generated by PROMALS3D. Each sequence (name shown in magenta color) is colored according to PSIPRED (secondary structure prediction tool: red, alpha-helix; blue, beta-strand). The target sequence below the templates is aligned based on predicted secondary structure and sequence similarity. Amino acid position is indicated in the beginning and end of each sequence. For the β-subunit, the alignment is shown from residue Cys143 which is numbered as 1 in the alignment. The first line in each block shows conservation indices only for positions with a value above five. The last two lines show consensus amino acid sequence (consensus_aa) and consensus predicted secondary structure (consensus_ss), respectively. Conserved amino acids are in bold and uppercase letters. Consensus amino acid symbols are: l, aliphatic (I, V, and L); @, aromatic (Y, H, W, and F); h, hydrophobic (W, F, Y, M, L, I, V, A, C, T, and H); o, alcohol (S and T); p, polar residues (D, E, H, K, N, Q, R, S, and T); t, tiny (A, G, C, and S); s, small (A, G, C, S, V, N, D, T, and P); b, bulky residues (E, F, I, K, L, M, Q, R, W, and Y); +, positively charged (K, R, and H); −, negatively charged (D and E); c, charged (D, E, K, R, and H). Consensus predicted secondary structure symbols are: h, alpha-helix; e, beta-strand. These alignments were used for modeling the alpha and beta subunit of acid ceramidase.
cge12316-sup-0003-FigureS3.epsPS document4286KFig. S3. Identification of disease causing mutations in Farber lipogranulomatosis patients. For each mutation, electropherogram showing the mutant sequence is on the left and for the normal sequence is on the right. Each mutation is indicated by an arrow in the respective electropherogram for the mutant sequence except for c.383-16delTTTTC which is indicated by a horizontal bar above the deleted sequence in the electropherogram for the normal sequence. The electropherogram for the c.383-16delTTTTC mutation is generated from mutant sequence cloned into TA cloning vector.
cge12316-sup-0004-FigureS4.epsPS document28233KFig. S4. Structure analysis of L182V mutation and of the G235 residue. In panel (a) model is shown in yellow surface with Leu182 in magenta and the closely located neighboring residue Asn173 (which is a site for glycosylation) in green. The inset shows no significant change in the interactions when Leu182Val substitution occurs. Panel (b) shows location of Gly235 in the glycine rich segment of the acid ceramidase β-subunit.
cge12316-sup-0005-TableS1.docWord document36KTable S1. List of primers used in the study
cge12316-sup-0006-AppendixS1.docWord document75KAppendix S1. Patient clinical details and additional methods.

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