title: Cloning a rat meprin cDNA reveals the enzyme is a heterodimer.
abstract: The structure of the kidney microvillar membrane metallopeptidase meprin (EC 3.4.24.18) from rats has been examined. Previously reported to be a homotetramer, we demonstrate that the enzyme is composed of two similar but distinct subunits through tryptic peptide mapping and the sequencing of peptides of the papain solubilized form of the enzyme. Two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis reveals that the native rat meprin tetramer is dissociated by detergent into disulfide-linked heterodimers. A full-length cDNA clone encoding one of the meprin subunits has been isolated and sequenced. The cDNA contains an open reading frame of 668 amino acids, coding for a polypeptide of molecular weight 75,054. The enzyme contains the zinc binding sequence HEFLH and a potential membrane-spanning region near its amino terminus. Comparison of this clone with peptide sequences from mouse meprins A and B shows that the clone is a B type or beta subunit. Northern blot analysis is consistent with the existence of two distinct subunits and further indicates that rat meprin subunits may be differentially expressed in various rat tissues.
authors.affiliation: Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas 75235-9038.
authors.forename: G D
authors.initials: GD
authors.lastname: Johnson
authors.forename: L B
authors.initials: LB
authors.lastname: Hersh
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D007668:Kidney / Q000201:enzymology; D008666:Metalloendopeptidases / Q000235:genetics*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010449:Peptide Mapping; D012313:RNA / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D014357:Trypsin
chemical_list: D012313:RNA; D004247:DNA; D014357:Trypsin; D008666:Metalloendopeptidases; C053833:meprin A