title: Krabbe disease: isolation and characterization of a full-length cDNA for human galactocerebrosidase.
abstract: Human galactocerebrosidase, the enzyme deficient in Krabbe disease, was purified, through several hydrophobic column steps and gel filtration, 22,650-fold from human lymphocytes. Using information on its N-terminal and internal amino acid sequences, and the polymerase chain reaction method, we cloned a full-length cDNA for the enzyme. The deduced amino acid sequence matched all amino acid sequences determined. The 3780 nucleotide sequence included 2007 nucleotides which encoded a single chain peptide of 669 amino acid residues with a 26 amino acid N-terminal signal peptide and six potential asparagine-linked glycosylation sites. The galactocerebrosidase cDNA detected an about 4 kb mRNA band material in human cultured skin fibroblasts. A nonsense mutation was found at codon 369 (GAA-->TAA) in the coding sequence of cDNA amplified from cultured skin fibroblast mRNA from a patient with typical Krabbe disease.
authors.affiliation: Department of Pediatrics, Osaka University School of Medicine, Japan.
authors.forename: N
authors.initials: N
authors.lastname: Sakai
authors.forename: K
authors.initials: K
authors.lastname: Inui
authors.forename: N
authors.initials: N
authors.lastname: Fujii
authors.forename: H
authors.initials: H
authors.lastname: Fukushima
authors.forename: J
authors.initials: J
authors.lastname: Nishimoto
authors.forename: I
authors.initials: I
authors.lastname: Yanagihara
authors.forename: Y
authors.initials: Y
authors.lastname: Isegawa
authors.forename: A
authors.initials: A
authors.lastname: Iwamatsu
authors.forename: S
authors.initials: S
authors.lastname: Okada
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005347:Fibroblasts / Q000737:chemistry; D005698:Galactosylceramidase / Q000172:deficiency / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D007965:Leukodystrophy, Globoid Cell / Q000201:enzymology / Q000235:genetics*; D008214:Lymphocytes / Q000201:enzymology; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D011498:Protein Precursors / Q000235:genetics; D021382:Protein Sorting Signals / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D017422:Sequence Analysis, DNA; D012867:Skin / Q000737:chemistry / Q000166:cytology
chemical_list: D018076:DNA, Complementary; D011498:Protein Precursors; D021382:Protein Sorting Signals; D012333:RNA, Messenger; D005698:Galactosylceramidase