title: Molecular cloning and characterization of bovine beta-mannosidase.
abstract: Deficiency of lysosomal beta-mannosidase activity results in a severe neurodegenerative disease in goats and cattle and a relatively milder phenotype in humans. A cDNA coding for the entire beta-mannosidase protein is described. Mixed oligonucleotides derived from bovine beta-mannosidase peptide sequences were used to screen a bovine thyroid cDNA library. Clones covering about 80% of the C-terminal region were recovered. The missing 5'-region was obtained using the technique of 5'-rapid amplification of cDNA ends. The composite cDNA contains 3852 nucleotides, encoding 879 amino acids. The N-terminal methionine is followed by 16 amino acids displaying the characteristics of a typical signal peptide sequence. The deduced amino acid sequence is colinear with all peptide sequences determined by protein microsequencing. Northern blot analysis demonstrates a single 4.2-kilobase transcript in various tissues from both normal and affected goats and calves. The mRNA level is decreased in tissues of affected beta-mannosidosis animals. The gene encoding beta-mannosidase is localized to human chromosome 4 as shown by Southern analysis of rodent/human somatic cell hybrids. This is the first report of cloning of lysosomal beta-mannosidase.
authors.affiliation: Department of Pathology, Michigan State University, East Lansing 48824.
authors.forename: H
authors.initials: H
authors.lastname: Chen
authors.forename: J R
authors.initials: JR
authors.lastname: Leipprandt
authors.forename: C E
authors.initials: CE
authors.lastname: Traviss
authors.forename: B L
authors.initials: BL
authors.lastname: Sopher
authors.forename: M Z
authors.initials: MZ
authors.lastname: Jones
authors.forename: K T
authors.initials: KT
authors.lastname: Cavanagh
authors.forename: K H
authors.initials: KH
authors.lastname: Friderici
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002417:Cattle; D003001:Cloning, Molecular; D018076:DNA, Complementary; D008247:Lysosomes / Q000201:enzymology; D008361:Mannosidases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D044902:beta-Mannosidase
chemical_list: D018076:DNA, Complementary; D008361:Mannosidases; D044902:beta-Mannosidase