title: Missense polymorphism in the human carboxypeptidase E gene alters enzymatic activity.
abstract: Carboxypeptidase E (CPE) is involved in the biosynthesis of peptide hormones and neurotransmitters, including insulin. One of the features of type 2 diabetes mellitus (T2DM) is an elevation in the proinsulin level and/or proinsulin/insulin molar ratio, suggesting that mutations in proinsulin processing enzymes may contribute to the development of T2DM. We scanned CPE for mutations in a collection of Ashkenazi T2DM families and identified five novel single nucleotide polymorphisms (SNPs). An SNP in the 283(rd) codon, c.847C>T, changes arginine to tryptophan (R283W). The residue Arg283 is conserved among CPE orthologs as well as most enzymatically active metallocarboxypeptidases. Of the 272 Ashkenazi T2DM pedigrees screened, we found four families segregating R283W. Within these four families, patients who inherited one copy of this variant had much earlier age of onset for T2DM. The R283W CPE protein cleaves peptide substrates with substantially lower efficiencies and is less stable at elevated temperature. In addition, the R283W CPE variant has a narrower pH optimum and is much less active at pH 6.0-6.5, indicating that the R283W CPE variant would be substantially less active than wild type CPE in the trans-Golgi network and immature secretory vesicles where the enzyme functions in vivo. To summarize, we uncovered a rare non-conservative missense mutation in CPE and demonstrated that the mutant protein has altered enzymatic properties. We predict that this mutant could cause hyperproinsulinism and diabetes in the homozygous state.
authors.affiliation: Millennium Pharmaceuticals, Inc., Cambridge, Massachusetts 02139, USA. chen@mpi.com
authors.forename: H
authors.initials: H
authors.lastname: Chen
authors.forename: S
authors.initials: S
authors.lastname: Jawahar
authors.forename: Y
authors.initials: Y
authors.lastname: Qian
authors.forename: Q
authors.initials: Q
authors.lastname: Duong
authors.forename: G
authors.initials: G
authors.lastname: Chan
authors.forename: A
authors.initials: A
authors.lastname: Parker
authors.forename: J M
authors.initials: JM
authors.lastname: Meyer
authors.forename: K J
authors.initials: KJ
authors.lastname: Moore
authors.forename: S
authors.initials: S
authors.lastname: Chayen
authors.forename: D J
authors.initials: DJ
authors.lastname: Gross
authors.forename: B
authors.initials: B
authors.lastname: Glaser
authors.forename: M A
authors.initials: MA
authors.lastname: Permutt
authors.forename: L D
authors.initials: LD
authors.lastname: Fricker
mesh_terms: D020121:5' Untranslated Regions / Q000235:genetics; D000595:Amino Acid Sequence; D043423:Carboxypeptidase H; D002268:Carboxypeptidases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D004252:DNA Mutational Analysis; D004795:Enzyme Stability; D005091:Exons / Q000235:genetics; D005260:Female; D005820:Genetic Testing; D006579:Heterozygote; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007585:Jews / Q000235:genetics; D007700:Kinetics; D008297:Male; D008969:Molecular Sequence Data; D020125:Mutation, Missense / Q000235:genetics*; D010375:Pedigree; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D016415:Sequence Alignment; D013696:Temperature; D044465:White People / Q000235:genetics
chemical_list: D020121:5' Untranslated Regions; D002268:Carboxypeptidases; D043423:Carboxypeptidase H