title: Functional consequences of PRODH missense mutations.
abstract: PRODH maps to 22q11 in the region deleted in the velocardiofacial syndrome/DiGeorge syndrome (VCFS/DGS) and encodes proline oxidase (POX), a mitochondrial inner-membrane enzyme that catalyzes the first step in the proline degradation pathway. At least 16 PRODH missense mutations have been identified in studies of type I hyperprolinemia (HPI) and schizophrenia, 10 of which are present at polymorphic frequencies. The functional consequences of these missense mutations have been inferred by evolutionary conservation, but none have been tested directly. Here, we report the effects of these mutations on POX activity. We find that four alleles (R185Q, L289M, A455S, and A472T) result in mild (<30%), six (Q19P, A167V, R185W, D426N, V427M, and R431H) in moderate (30%-70%), and five (P406L, L441P, R453C, T466M, and Q521E) in severe (>70%) reduction in POX activity, whereas one (Q521R) increases POX activity. The POX encoded by one severe allele (T466M) shows in vitro responsiveness to high cofactor (flavin adenine dinucleotide) concentrations. Although there is limited information on plasma proline levels in individuals of known PRODH genotype, extant data suggest that severe hyperprolinemia (>800 microM) occurs in individuals with large deletions and/or PRODH missense mutations with the most-severe effect on function (L441P and R453C), whereas modest hyperprolinemia (300-500 microM) is associated with PRODH alleles with a moderate reduction in activity. Interestingly, three of the four alleles associated with or found in schizophrenia (V427M, L441P, and R453C) resulted in severe reduction of POX activity and hyperprolinemia. These observations plus the high degree of polymorphism at the PRODH locus are consistent with the hypothesis that reduction in POX function is a risk factor for schizophrenia.
authors.affiliation: Howard Hughes Medical Institute, and Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
authors.forename: Hans-Ulrich
authors.initials: HU
authors.lastname: Bender
authors.forename: Shlomo
authors.initials: S
authors.lastname: Almashanu
authors.forename: Gary
authors.initials: G
authors.lastname: Steel
authors.forename: Chien-An
authors.initials: CA
authors.lastname: Hu
authors.forename: Wei-Wen
authors.initials: WW
authors.lastname: Lin
authors.forename: Alecia
authors.initials: A
authors.lastname: Willis
authors.forename: Ann
authors.initials: A
authors.lastname: Pulver
authors.forename: David
authors.initials: D
authors.lastname: Valle
mesh_terms: D000483:Alleles; D000595:Amino Acid Sequence; D020134:Catalytic Domain / Q000235:genetics; D003001:Cloning, Molecular; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D020125:Mutation, Missense; D010641:Phenotype; D011392:Proline / Q000097:blood; D011394:Proline Oxidase / Q000737:chemistry / Q000172:deficiency / Q000235:genetics* / Q000502:physiology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012559:Schizophrenia / Q000201:enzymology / Q000235:genetics; D017386:Sequence Homology, Amino Acid
chemical_list: D011994:Recombinant Proteins; D005182:Flavin-Adenine Dinucleotide; D011392:Proline; D011394:Proline Oxidase