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Peroxisomal fatty acid oxidation disorders and 58 kDa sterol carrier protein X (SCPx). Activity measurements in liver and fibroblasts using a newly developed method.
J Lipid Res. 2000 Mar; 41(3):336-42.JL

Abstract

Sterol carrier protein X (SCPx) plays a crucial role in the peroxisomal oxidation of branched-chain fatty acids. To investigate whether patients with an unresolved defect in peroxisomal beta-oxidation are deficient for SCPx, we developed a novel and specific assay to measure the activity of SCPx in both liver and fibroblast homogenates. The substrate used in the assay, 3alpha, 7alpha,12alpha-trihydroxy-24-keto-5beta-cholestanoy l-CoA (24-keto-THC-CoA), is produced by preincubating the enoyl-CoA of the bile acid intermediate THCA with a lysate from the yeast Saccharomyces cerevisiae expressing human D-bifunctional protein. After the preincubation period, liver or fibroblast homogenate is added plus CoASH, and the production of choloyl-CoA is determined by HPLC. The specificity of the assay was demonstrated by the finding of a full deficiency in fibroblasts from an SCPx knock-out mouse. In addition to SCPx activity measurements in fibroblasts from patients with a defect in peroxisomal beta-oxidation of unresolved etiology, we studied the stability and activity of SCPx in fibroblasts from patients with Zellweger syndrome, which lack functional peroxisomes. We found that SCPx is not only stable in the cytosol, but displays a higher activity in fibroblasts from patients with Zellweger syndrome than in control fibroblasts. Furthermore, in all patients studied with a defect in peroxisomal beta-oxidation of unknown origin, SCPx was found to be normally active, indicating that human SCPx deficiency remains to be identified.

Authors+Show Affiliations

Department of Clinical Chemistry, Academic Medical Center, University of Amsterdam, 1100 DE Amsterdam, The Netherlands.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

10706581

Citation

Ferdinandusse, S, et al. "Peroxisomal Fatty Acid Oxidation Disorders and 58 kDa Sterol Carrier Protein X (SCPx). Activity Measurements in Liver and Fibroblasts Using a Newly Developed Method." Journal of Lipid Research, vol. 41, no. 3, 2000, pp. 336-42.
Ferdinandusse S, Denis S, van Berkel E, et al. Peroxisomal fatty acid oxidation disorders and 58 kDa sterol carrier protein X (SCPx). Activity measurements in liver and fibroblasts using a newly developed method. J Lipid Res. 2000;41(3):336-42.
Ferdinandusse, S., Denis, S., van Berkel, E., Dacremont, G., & Wanders, R. J. (2000). Peroxisomal fatty acid oxidation disorders and 58 kDa sterol carrier protein X (SCPx). Activity measurements in liver and fibroblasts using a newly developed method. Journal of Lipid Research, 41(3), 336-42.
Ferdinandusse S, et al. Peroxisomal Fatty Acid Oxidation Disorders and 58 kDa Sterol Carrier Protein X (SCPx). Activity Measurements in Liver and Fibroblasts Using a Newly Developed Method. J Lipid Res. 2000;41(3):336-42. PubMed PMID: 10706581.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Peroxisomal fatty acid oxidation disorders and 58 kDa sterol carrier protein X (SCPx). Activity measurements in liver and fibroblasts using a newly developed method. AU - Ferdinandusse,S, AU - Denis,S, AU - van Berkel,E, AU - Dacremont,G, AU - Wanders,R J, PY - 2000/3/8/pubmed PY - 2000/5/16/medline PY - 2000/3/8/entrez SP - 336 EP - 42 JF - Journal of lipid research JO - J Lipid Res VL - 41 IS - 3 N2 - Sterol carrier protein X (SCPx) plays a crucial role in the peroxisomal oxidation of branched-chain fatty acids. To investigate whether patients with an unresolved defect in peroxisomal beta-oxidation are deficient for SCPx, we developed a novel and specific assay to measure the activity of SCPx in both liver and fibroblast homogenates. The substrate used in the assay, 3alpha, 7alpha,12alpha-trihydroxy-24-keto-5beta-cholestanoy l-CoA (24-keto-THC-CoA), is produced by preincubating the enoyl-CoA of the bile acid intermediate THCA with a lysate from the yeast Saccharomyces cerevisiae expressing human D-bifunctional protein. After the preincubation period, liver or fibroblast homogenate is added plus CoASH, and the production of choloyl-CoA is determined by HPLC. The specificity of the assay was demonstrated by the finding of a full deficiency in fibroblasts from an SCPx knock-out mouse. In addition to SCPx activity measurements in fibroblasts from patients with a defect in peroxisomal beta-oxidation of unresolved etiology, we studied the stability and activity of SCPx in fibroblasts from patients with Zellweger syndrome, which lack functional peroxisomes. We found that SCPx is not only stable in the cytosol, but displays a higher activity in fibroblasts from patients with Zellweger syndrome than in control fibroblasts. Furthermore, in all patients studied with a defect in peroxisomal beta-oxidation of unknown origin, SCPx was found to be normally active, indicating that human SCPx deficiency remains to be identified. SN - 0022-2275 UR - https://www.unboundmedicine.com/medline/citation/10706581/Peroxisomal_fatty_acid_oxidation_disorders_and_58_kDa_sterol_carrier_protein_X__SCPx___Activity_measurements_in_liver_and_fibroblasts_using_a_newly_developed_method_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-2275(20)34472-2 DB - PRIME DP - Unbound Medicine ER -