title: alpha-Mannosidosis: functional cloning of the lysosomal alpha-mannosidase cDNA and identification of a mutation in two affected siblings.
abstract: a-Mannosidosis (MIM 248500) is an autosomal recessive lysosomal storage disorder resulting from deficient activity of lysosomal alpha-mannosidase (LAMAN) (EC 3.2.1.24). The disease is characterized by massive intracellular accumulation of mannose-rich oligosaccharides with resulting mental retardation, hearing loss, immune deficiency and skeletal changes. We report here the purification and characterization of human placenta LAMAN. The enzyme is synthesized as a single-chain precursor which is processed into three glycopeptides of 70, 42 and 15 kDa. The 70 kDa peptide is further partially proteolysed into three more peptides that are joined by disulfide bridges. The laman cDNA sequence was assembled from overlapping fragments obtained by PCR on human fibroblast and human lung cDNA. The deduced amino acid sequence contains a putative signal peptide of 48 amino acids followed by a polypeptide sequence of 962 amino acids. Northern blot analyses revealed a single transcript of approximately 3.5 kb present in all tissues examined but at varying levels. Two affected siblings of Palestinian origin were homozygous for a mutation that causes a His-->Leu replacement at a position which is conserved among class 2 alpha-mannosidases from several species.
authors.affiliation: Department of Medical Genetics, University Hospital and University of Tromso, Norway. oivindn@fagmed.uit.no
authors.forename: O
authors.initials: O
authors.lastname: Nilssen
authors.forename: T
authors.initials: T
authors.lastname: Berg
authors.forename: H M
authors.initials: HM
authors.lastname: Riise
authors.forename: U
authors.initials: U
authors.lastname: Ramachandran
authors.forename: G
authors.initials: G
authors.lastname: Evjen
authors.forename: G M
authors.initials: GM
authors.lastname: Hansen
authors.forename: D
authors.initials: D
authors.lastname: Malm
authors.forename: L
authors.initials: L
authors.lastname: Tranebjaerg
authors.forename: O K
authors.initials: OK
authors.lastname: Tollersrud
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002417:Cattle; D003001:Cloning, Molecular; D017124:Conserved Sequence; D003429:Cross Reactions; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D006020:Glycopeptides / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006801:Humans; D008247:Lysosomes / Q000201:enzymology*; D008297:Male; D008361:Mannosidases / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D010920:Placenta / Q000737:chemistry / Q000201:enzymology; D011247:Pregnancy; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution; D043323:alpha-Mannosidase; D008363:alpha-Mannosidosis / Q000235:genetics*
chemical_list: D018076:DNA, Complementary; D006020:Glycopeptides; D010446:Peptide Fragments; D011498:Protein Precursors; D008361:Mannosidases; D043323:alpha-Mannosidase