title: Measuring propionyl-CoA carboxylase activity in phytohemagglutinin stimulated lymphocytes using high performance liquid chromatography.
abstract: BACKGROUND
  Propionyl-CoA carboxylase (PCC) is a mitochondrial enzyme involved in the catabolism of several essential amino acids and odd chain fatty acids. Previous PCC assays have involved either a radiometric assay or have required mitochondria isolation and/or enzyme purification.
  
  
  METHODS
  We developed an enzymatic method to analyze PCC activity in phytohemagglutinin (PHA) stimulated lymphocytes that involves high performance liquid chromatography.
  
  
  RESULTS
  The method shows good linearity and sensitivity. PCC activity was unaffected even when lymphocytes were isolated and PHA stimulated after a whole blood sample had been stored at 4°C for 5days. This indicates that this method is suitable for analyzing samples from distant medical centers. The PCC activity of patients with propionic acidemia was found to be much lower than that of normal individuals and carriers. However, this PCC assay is significantly affected by the red blood cell contamination. In conclusion, this is a reliable method for performing PCC assays and only requires 0.5 to 1.0ml of whole blood from newborns.
  
  
  CONCLUSIONS
  The PCC assay established in this study is useful for the confirmation of PA in individuals, and prenatal diagnosis and genetic counseling for the affected families.
authors.affiliation: Department of Education and Research, Taipei City Hospital, Taipei, Taiwan.
authors.forename: Yu-Ning
authors.initials: YN
authors.lastname: Liu
authors.affiliation: Department of Education and Research, Taipei City Hospital, Taipei, Taiwan; VYM Genome Research Center, National Yang-Ming University, Taipei, Taiwan.
authors.forename: Tze-Tze
authors.initials: TT
authors.lastname: Liu
authors.affiliation: Department of Education and Research, Taipei City Hospital, Taipei, Taiwan.
authors.forename: Ya-Ling
authors.initials: YL
authors.lastname: Fan
authors.affiliation: Department of Pediatrics, Taipei Veterans General Hospital, Taipei, Taiwan.
authors.forename: Dau-Ming
authors.initials: DM
authors.lastname: Niu
authors.affiliation: Department of Medical Genetics, National Taiwan University Hospital, Taipei, Taiwan.
authors.forename: Yin-Hsiu
authors.initials: YH
authors.lastname: Chien
authors.affiliation: Department of Pediatrics, National Cheng Kung University Hospital, Tainan, Taiwan.
authors.forename: Yen-Yin
authors.initials: YY
authors.lastname: Chou
authors.affiliation: Department of Medical Genetics, National Taiwan University Hospital, Taipei, Taiwan.
authors.forename: Ni-Chung
authors.initials: NC
authors.lastname: Lee
authors.affiliation: Department of Education and Research, Taipei City Hospital, Taipei, Taiwan; Preventive Medicine Foundations, Taipei, Taiwan.
authors.forename: Kwang-Jen
authors.initials: KJ
authors.lastname: Hsiao
authors.affiliation: Department of Education and Research, Taipei City Hospital, Taipei, Taiwan. Electronic address: yhchiu@ntu.edu.tw.
authors.forename: Yen-Hui
authors.initials: YH
authors.lastname: Chiu
mesh_terms: D000293:Adolescent; D002648:Child; D002851:Chromatography, High Pressure Liquid; D057075:Enzyme Assays / Q000379:methods*; D004795:Enzyme Stability; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D057230:Limit of Detection; D016014:Linear Models; D008214:Lymphocytes / Q000187:drug effects* / Q000201:enzymology*; D008297:Male; D043586:Methylmalonyl-CoA Decarboxylase / Q000737:chemistry / Q000378:metabolism*; D010835:Phytohemagglutinins / Q000494:pharmacology*
chemical_list: D010835:Phytohemagglutinins; D043586:Methylmalonyl-CoA Decarboxylase
keywords: Propionic acidemia (PA); Propionyl-CoA carboxylase (PCC)