title: Identification and characterization of temperature-sensitive mild mutations in three Japanese patients with nonsevere forms of very-long-chain acyl-CoA dehydrogenase deficiency.
abstract: Very-long-chain acyl-CoA dehydrogenase (VLCAD) deficiency is clinically classified into severe, intermediate, and myopathic forms. We identified mutations in three unrelated Japanese patients with VLCAD deficiency: two with the myopathic form and one with the intermediate form, all compound heterozygotes of K264E/M437V, A416T/1798delA, and P89S/IVS16-3delAA, respectively. We characterized four missense mutations, K264E, M437V, A416T, and P89S, by transisent expression analysis, using SV40-transformed fibroblasts derived from a VLCAD-null patient, as recipient cells. In transient expression of the wild-type VLCAD cDNA, VLCAD activity at 30 degrees C was higher than at 37 degrees C. Moreover, this temperature-sensitive character is more evident in all the mutant proteins tested than in wild type. Based on characterization of the five missense mutations identified in four Japanese patients, including data on one patient with the myopathic form previously reported, patients with the nonsevere forms (intermediate or myopathic forms) have missense mutations with residual activities in at least one allele. Expression analysis at 30 degrees C may be more useful for evaluating these missense mutations, compared with that at 37 degrees C.
authors.affiliation: Department of Pediatrics, Shimane Medical University, Izumo, Shimane 693-8501, Japan. mysunao@shimane-med.ac.jp
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Takusa
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Fukao
authors.forename: Masahiko
authors.initials: M
authors.lastname: Kimura
authors.forename: Atsushi
authors.initials: A
authors.lastname: Uchiyama
authors.forename: Wataru
authors.initials: W
authors.lastname: Abo
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Tsuboi
authors.forename: Shinichi
authors.initials: S
authors.lastname: Hirose
authors.forename: Hideki
authors.initials: H
authors.lastname: Fujioka
authors.forename: Naomi
authors.initials: N
authors.lastname: Kondo
authors.forename: Seiji
authors.initials: S
authors.lastname: Yamaguchi
mesh_terms: D044942:Acyl-CoA Dehydrogenase, Long-Chain; D044944:Acyl-CoA Dehydrogenases / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism; D000328:Adult; D002461:Cell Line, Transformed; D002648:Child; D004252:DNA Mutational Analysis; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology; D006801:Humans; D015151:Immunoblotting; D007564:Japan; D008297:Male; D009135:Muscular Diseases / Q000201:enzymology / Q000235:genetics* / Q000473:pathology; D009154:Mutation; D012867:Skin / Q000166:cytology / Q000201:enzymology; D013696:Temperature
chemical_list: D018076:DNA, Complementary; D044944:Acyl-CoA Dehydrogenases; D044942:Acyl-CoA Dehydrogenase, Long-Chain