title: Cloning of a human acid sphingomyelinase cDNA with a new mutation that renders the enzyme inactive.
abstract: A cDNA encoding human acid sphingomyelinase was initially obtained by screening a placental cDNA library in lambda gt11 with a synthetic oligonucleotide probe and subsequently with partial cDNA. The full-length cDNA, hPSM55, comprised 2,376 nucleotides, with a 5' untranslated sequence of 122 nucleotides, an open reading frame of 1,884 nucleotides encoding a protein of 627 amino acids, and a 3' untranslated region of 370 bases. hPSM55 was almost identical to pASM-1FL reported by Schuchman et al. (J. Biol. Chem. 266, 8531-8539, 1991) except for a 6 base pair deletion in the signal peptide, which indicated the possible removal of valine and leucine residues between positions 36 and 37, and a 463T- to C-transition, which indicated a possible substitution of 155arginine for cystine. This cDNA was expressed in both COS-7 cells and Chinese hamster ovary cells. There was no increase in acid sphingomyelinase activity in either cell line following transfection. However, the correction of a single base change, 463C to T, in hPSM55 caused increased acid sphingomyelinase activity in transfectants. These results suggest that the mutation of nucleotide 463C to T plays an important role in the catalytic activity of acid sphingomyelinase.
authors.affiliation: Department of Pediatrics, Georgetown University Medical Center, Washington, DC.
authors.forename: H
authors.initials: H
authors.lastname: Ida
authors.forename: O M
authors.initials: OM
authors.lastname: Rennert
authors.forename: Y
authors.initials: Y
authors.lastname: Eto
authors.forename: W Y
authors.initials: WY
authors.lastname: Chan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D016466:CHO Cells; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D015345:Oligonucleotide Probes; D010920:Placenta / Q000737:chemistry; D021382:Protein Sorting Signals / Q000737:chemistry; D013108:Sphingomyelin Phosphodiesterase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: D018076:DNA, Complementary; D015345:Oligonucleotide Probes; D021382:Protein Sorting Signals; D013108:Sphingomyelin Phosphodiesterase