caa a22 4500 397574142 CHVBK 20180308164850.0 cr unu---uuuuu 161202e199604 xx s 000 0 eng 10.1093/hmg/5.4.461 doi (NATIONALLICENCE)oxford-10.1093/hmg/5.4.461 Cloning and Characterization of Human Very-Long-Chain Acyl-CoA Dehydrogenase cDNA, Chromosomal Assignment of the Gene and Identification in Four Patients of Nine Different Mutations Within the VLCAD Gene [Elektronische Daten] [Brage Storstein Andresen, Peter Bross, Christine Vianey-Saban, Priscille Divry, Marie-Thérèse Zabot, Charles R. Roe, Mohamed A. Nada, Annette Byskov, Torben A. Kruse, Søren Neve, Karsten Kristiansen, Inga Knudsen, Morten Juhl Corydon, Niels Gregersen] Very-long-chain acyl-CoA dehydrogenase (VLCAD) is one of four straight-chain acyl-CoA dehydrogenase (ACD) enzymes, which are all nuclear encoded mitochondrial flavoproteins catalyzing the initial step in fatty acid β-oxidation. We have used the very fast, Rapid Amplification of cDNA Ends (RACE) based strategy to obtain the sequence of cDNAs encoding human VLCAD from placenta and fibroblasts. Alignment of the predicted amino acid sequence of human VLCAD with those of the other human ACD enzymes revealed extensive sequence homology. Moreover, human VLCAD and human acyl-CoA oxidase showed extensive sequence homology corroborating the notion that these genes are evolutionarily related. Southern blot analysis of genomic DNA from hybrid cell lines was used to localize the VLCAD gene to human chromosome 17p11.2-p11.13105. Using Northern and Western blot analysis to investigate the tissue specific distribution of VLCAD mRNA and protein in several human tissues we showed that VLCAD is most abundant in heart and skeletal muscle. This agrees well with the fact that cardiac and muscle symptoms are characteristic for patients with VLCAD deficiency. Northern blot analysis and sequencing of cloned PCR amplified VLCAD cDNA from four unrelated patients with VLCAD deficiency showed that VLCAD mRNA was undetectable in one patient and that the other three have mutations in both VLCAD alleles. Western blot analysis of patient fibroblasts showed that the identified mutations result in severely reduced amounts of VLCAD protein. None of the patients harbored identical mutations suggesting that the mutational heterogeneity in VLCAD deficiency is large. © 1996 Oxford University Press Andresen Brage Storstein Center for Medical Molecular Biology, Aarhus University Hospital and Faculty of Health Science, Skejby Sygehus, Aarhus, Denmark aut Bross Peter Center for Medical Molecular Biology, Aarhus University Hospital and Faculty of Health Science, Skejby Sygehus, Aarhus, Denmark aut Vianey-Saban Christine Unité d'Etude des Maladies Métaboliques, Hôpital Debrousse, Lyon, France aut Divry Priscille Unité d'Etude des Maladies Métaboliques, Hôpital Debrousse, Lyon, France aut Zabot Marie-Thérèse Unité d'Etude des Maladies Métaboliques, Hôpital Debrousse, Lyon, France aut Roe Charles R. Institute of Metabolic Disease, Baylor University Medical Center, Dallas, Texas, USA aut Nada Mohamed A. Institute of Metabolic Disease, Baylor University Medical Center, Dallas, Texas, USA aut Byskov Annette Danish Center for Human Genome Research, Aarhus, Denmark aut Kruse Torben A. Danish Center for Human Genome Research, Aarhus, Denmark aut Neve Søren Department of Molecular Biology, University of Odense, Odense, Denmark aut Kristiansen Karsten Department of Molecular Biology, University of Odense, Odense, Denmark aut Knudsen Inga Center for Medical Molecular Biology, Aarhus University Hospital and Faculty of Health Science, Skejby Sygehus, Aarhus, Denmark aut Corydon Morten Juhl Center for Medical Molecular Biology, Aarhus University Hospital and Faculty of Health Science, Skejby Sygehus, Aarhus, Denmark aut Gregersen Niels Center for Medical Molecular Biology, Aarhus University Hospital and Faculty of Health Science, Skejby Sygehus, Aarhus, Denmark aut Human Molecular Genetics Oxford University Press 5/4(1996-04), 461-472 0964-6906 5:4<461 1996 5 hmg https://doi.org/10.1093/hmg/5.4.461 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/hmg/5.4.461 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Andresen Brage Storstein Center for Medical Molecular Biology, Aarhus University Hospital and Faculty of Health Science, Skejby Sygehus, Aarhus, Denmark aut NATIONALLICENCE 700 1- Bross Peter Center for Medical Molecular Biology, Aarhus University Hospital and Faculty of Health Science, Skejby Sygehus, Aarhus, Denmark aut NATIONALLICENCE 700 1- Vianey-Saban Christine Unité d'Etude des Maladies Métaboliques, Hôpital Debrousse, Lyon, France aut NATIONALLICENCE 700 1- Divry Priscille Unité d'Etude des Maladies Métaboliques, Hôpital Debrousse, Lyon, France aut NATIONALLICENCE 700 1- Zabot Marie-Thérèse Unité d'Etude des Maladies Métaboliques, Hôpital Debrousse, Lyon, France aut NATIONALLICENCE 700 1- Roe Charles R. Institute of Metabolic Disease, Baylor University Medical Center, Dallas, Texas, USA aut NATIONALLICENCE 700 1- Nada Mohamed A. Institute of Metabolic Disease, Baylor University Medical Center, Dallas, Texas, USA aut NATIONALLICENCE 700 1- Byskov Annette Danish Center for Human Genome Research, Aarhus, Denmark aut NATIONALLICENCE 700 1- Kruse Torben A. Danish Center for Human Genome Research, Aarhus, Denmark aut NATIONALLICENCE 700 1- Neve Søren Department of Molecular Biology, University of Odense, Odense, Denmark aut NATIONALLICENCE 700 1- Kristiansen Karsten Department of Molecular Biology, University of Odense, Odense, Denmark aut NATIONALLICENCE 700 1- Knudsen Inga Center for Medical Molecular Biology, Aarhus University Hospital and Faculty of Health Science, Skejby Sygehus, Aarhus, Denmark aut NATIONALLICENCE 700 1- Corydon Morten Juhl Center for Medical Molecular Biology, Aarhus University Hospital and Faculty of Health Science, Skejby Sygehus, Aarhus, Denmark aut NATIONALLICENCE 700 1- Gregersen Niels Center for Medical Molecular Biology, Aarhus University Hospital and Faculty of Health Science, Skejby Sygehus, Aarhus, Denmark aut NATIONALLICENCE 773 0- Human Molecular Genetics Oxford University Press 5/4(1996-04), 461-472 0964-6906 5:4<461 1996 5 hmg Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford