Molecular Cloning and Nucleotide Sequence of Complementary DNA for Human Hepatic Cytosolic Acetoacetyl-Coenzyme A Thiolase

Song, Xiaolin; Fukao, Toshiyuki; Yamaguchi, Shun; Miyazawa, Shintaro; Hashimoto, T.; Orii, T

doi:10.1006/bbrc.1994.1726

Cited by 38 publications

(26 citation statements)

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“…A search of GenBank revealed that 22 of the insert sequences were cytosolic thiolase (EC 2.3.1.9), and 32 were the C-terminal domain of a protein we have called ␥-SNAP-associated factor-1 (Gaf-1). The clones recovered for cytosolic thiolase were full-length and identical to the clone previously reported by Song et al (24). Re-transformation assays showed that thiolase and Gaf-1 fusion proteins interact specifically with the ␥-SNAP bait because no interaction was detected with either Gal4-DBD alone or Gal4-DBD-p53 (data not shown).…”

Section: Identification Of ␥-Snap-bindingsupporting

confidence: 80%

Gaf-1, a γ-SNAP-binding Protein Associated with the Mitochondria

Chen

et al. 2001

Journal of Biological Chemistry

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The role of ␣/␤-SNAP (Soluble NSF Attachment Protein) in vesicular trafficking is well established; however, the function of the ubiquitously expressed ␥-SNAP remains unclear. To further characterize the cellular role of this enigmatic protein, a two-hybrid screen was used to identify new, ␥-SNAP-binding proteins and to uncover potentially novel functions for ␥-SNAP. One such SNAP-binding protein, termed Gaf-1 (␥-SNAP associate factor-1) specifically binds ␥-but not ␣-SNAP. The full-length Gaf-1 (75 kDa) is ubiquitously expressed and is found stoichiometrically associated with ␥-SNAP in cellular extracts. This binding is distinct from other SNAP interactions since no ␣-SNAP or NSF coprecipitated with Gaf-1. Subcellular fractionation and immunofluorescence analysis show that Gaf-1 is peripherally associated with the outer mitochondrial membrane. Only a fraction of ␥-SNAP was mitochondrial with the balance being either cytosolic or associated with other membrane fractions. GFP-␥-SNAP and the C-terminal domain of Gaf-1 both show a reticular distribution in HEK-293 cells. This reticular structure colocalizes with Gaf-1 and mitochondria as well as with microtubules but not with other cytoskeletal elements. These data identify a class of ␥-SNAP interactions that is distinct from other members of the SNAP family and point to a potential role for ␥-SNAP in mitochondrial dynamics.The soluble N-ethylmaleimide-sensitive factor (NSF) 1 attachment protein family (␣-, ␤-, and ␥-SNAP) is thought to function in vesicular transport by mediating the binding of the NSF to the SNAP receptors (SNAREs) (reviewed in Refs.

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Section: Identification Of ␥-Snap-bindingsupporting

confidence: 80%

Gaf-1, a γ-SNAP-binding Protein Associated with the Mitochondria

Chen

et al. 2001

Journal of Biological Chemistry

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“…Wild-type and mutant SCOT expression vectors (4 g) were transfected using Lipofectamine 2000 (Invitrogen, Carlsbad, CA) in ‫01ف‬ 5 SV40-transformed SCOT-deficient fibroblasts from patient GS02, which contain no endogenous SCOT activity (18). One microgram of the cytosolic acetoacetyl-CoA thiolase (CT)-expressing vector, pCAGGSct (20), was co-transfected to monitor transfection efficiency. After a 72-h incubation at 30°C, 37°C, or 39.5°C, cells were harvested and stored at Ϫ80°C until the assay for SCOT (18) and CT (17) activities.…”

Section: Methodsmentioning

confidence: 99%

Patients Homozygous for the T435N Mutation of Succinyl-CoA:3-Ketoacid CoA Transferase (SCOT) Do Not Show Permanent Ketosis

Fukao

Shintaku²,

Kusubae

et al. 2004

Pediatr Res

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“…After a 72-h incubation period at 37°C or 30°C, the cells were harvested and assayed for C16-DH activity. Acetoacetyl-CoA thiolase activity was assayed, as described (17).…”

Section: Methodsmentioning

confidence: 99%

Myopathic Form of Very-Long Chain Acyl-CoA Dehydrogenase Deficiency: Evidence for Temperature-Sensitive Mild Mutations in Both Mutant Alleles in a Japanese Girl

et al. 2001

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In a 14-year-old Japanese girl, manifested recurrent myalgia with elevated serum creatine kinase after moderate exercise became evident, and she was diagnosed as having a myopathic form of very-long chain acyl-CoA dehydrogenase deficiency. Her first clinical symptom of the disease was evident when she was 6 y of age. She had never had hypoglycemic attacks, and hepatomegaly and cardiomyopathy were absent. The diagnosis was suspected on the basis of the urinary organic acid profile after a 36-h fast, long-chain fatty acid-loading test, and the blood acylcarnitine profile. Acyl-CoA dehydrogenase activity with palmitoyl-CoA as a substrate was severely decreased in her fibroblasts, and the amount of very-long chain acyl-CoA dehydrogenase protein was reduced. She was a compound heterozygote of A416T from her father and R450H from her mother. Transient expression of mutant A416T cDNA retained a significant residual acyl-CoA dehydrogenase activity of 10% and 20% normal at 37°C and 30°C, respectively. Specific activity of A416T mutant protein was calculated to be one fifth that of control. In the case of R450H mutant expression, a low residual acyl-CoA dehydrogenase activity of 5% normal was detected at 30°C although significant activity was absent at 37°C. The R450H protein was not detected at 37°C but was clearly detected at one fourth the normal amount at 30°C. These results indicate that both mutations were temperature-sensitive mild mutations, the result being the mildest phenotype of very-long chain acylCoA dehydrogenase deficiency. VLCAD(EC 1.3.99.13; HUGO symbol, ACADVL) belongs to the ␤-oxidation system of long-chain fatty acids together with mitochondrial enoyl-CoA hydratase/3-hydroxyacyl-CoA dehydrogenase/3-ketoacyl-CoA thiolase trifunctional protein (1, 2).The clinical phenotype of VLCAD deficiency (MIM# 201475) was first divided into two different forms (3); a severe presentation characterized by early onset of symptoms, with hypertrophic cardiomyopathy and a high incidence of death, and a milder form with a late onset with Reye-like disease, hypoketotic hypoglycemia, no cardiac involvement, and a more favorable outcome. Later occurrences in adolescents and adults have been reported, with the absence of hypoglycemia and cardiomyopathy but with episodes of rhabdomyolysis and myoglobinuria, usually triggered by exercise or fasting (4 -7). Hence VLCAD deficiency can be separated into three clinical phenotypes (8).Gene mutations in VLCAD deficiency have been reported, with a wide spectrum (8 -14). Andresen et al. (8) suggested a clear correlation of genotype with disease phenotype in VLCAD deficiency, from data on 58 independent mutations in 55 unrelated patients. They reported that most patients with the severe childhood form had two null mutations and that most

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Molecular Cloning and Nucleotide Sequence of Complementary DNA for Human Hepatic Cytosolic Acetoacetyl-Coenzyme A Thiolase

Cited by 38 publications

References 0 publications

Gaf-1, a γ-SNAP-binding Protein Associated with the Mitochondria

Gaf-1, a γ-SNAP-binding Protein Associated with the Mitochondria

Patients Homozygous for the T435N Mutation of Succinyl-CoA:3-Ketoacid CoA Transferase (SCOT) Do Not Show Permanent Ketosis

Myopathic Form of Very-Long Chain Acyl-CoA Dehydrogenase Deficiency: Evidence for Temperature-Sensitive Mild Mutations in Both Mutant Alleles in a Japanese Girl

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