PMID- 15698607
OWN - NLM
STAT- MEDLINE
DCOM- 20050422
LR  - 20131121
IS  - 0009-8981 (Print)
IS  - 0009-8981 (Linking)
VI  - 353
IP  - 1-2
DP  - 2005 Mar
TI  - Establishment of a practical enzymatic assay method for determination of 
      isovaleryl-CoA dehydrogenase activity using high-performance liquid 
      chromatography.
PG  - 193-9
AB  - BACKGROUND: Isovaleric acidemia (IVA) is one of the various target disorders for 
      tandem mass spectrometry (MS/MS) newborn screening. In the diagnosis of IVA, no 
      enzymatic assay method for isovaleryl-CoA dehydrogenase (IVD) activity has been 
      reported whereby the production of enoyl-CoA species was directly detected. We 
      established a direct assay method to detect 3-methylcrotonyl-CoA (MC-CoA) 
      production using high-performance liquid chromatography (HPLC). METHODS: 
      Isovaleryl-CoA dehydrogenase crude enzyme was prepared by sonicating lymphocytes 
      in peripheral blood. Aliquots were incubated with isovaleryl-CoA, flavin adenine 
      dinucleotide, and phenazine methosulfate. 3-Methylcrotonyl-CoA produced in the 
      samples was separated by HPLC and detected using an ultraviolet 
      spectrophotometer. RESULTS: The detection of MC-CoA was reproducible depending 
      upon the concentration of the substrates, the incubation time, and the number of 
      cells contained in the crude enzyme solution. We applied this assay to three 
      patients diagnosed with IVA and showed that neither of them had detectable 
      residual activity. Only a few hours were required from the initial blood sampling 
      to the end of the assay. CONCLUSIONS: These results demonstrate that this method 
      for detecting MC-CoA production, using HPLC, is a practical assay for determining 
      IVD activity. It can be a useful confirmatory test for IVA cases detected through 
      MS/MS screening of newborns.
FAU - Tajima, Go
AU  - Tajima G
AD  - Department of Pediatrics, Hiroshima University Graduate School of Biomedical 
      Sciences, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8551, Japan. ugoch@mac.com
FAU - Sakura, Nobuo
AU  - Sakura N
FAU - Yofune, Hiroko
AU  - Yofune H
FAU - Dwi Bahagia Febriani, Andi
AU  - Dwi Bahagia Febriani A
FAU - Nishimura, Yutaka
AU  - Nishimura Y
FAU - Sakamoto, Akiko
AU  - Sakamoto A
FAU - Ono, Hiroaki
AU  - Ono H
FAU - Shigematsu, Yosuke
AU  - Shigematsu Y
FAU - Kobayashi, Masao
AU  - Kobayashi M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - Clin Chim Acta
JT  - Clinica chimica acta; international journal of clinical chemistry
JID - 1302422
RN  - EC 1.3.- (Oxidoreductases Acting on CH-CH Group Donors)
RN  - EC 1.3.8.4 (Isovaleryl-CoA Dehydrogenase)
SB  - IM
MH  - Chromatography, High Pressure Liquid/*methods
MH  - Humans
MH  - Isovaleryl-CoA Dehydrogenase
MH  - Oxidoreductases Acting on CH-CH Group Donors/*metabolism
MH  - Reproducibility of Results
EDAT- 2005/02/09 09:00
MHDA- 2005/04/23 09:00
CRDT- 2005/02/09 09:00
PHST- 2004/07/19 00:00 [received]
PHST- 2004/11/07 00:00 [revised]
PHST- 2004/11/08 00:00 [accepted]
PHST- 2005/02/09 09:00 [pubmed]
PHST- 2005/04/23 09:00 [medline]
PHST- 2005/02/09 09:00 [entrez]
AID - S0009-8981(04)00546-7 [pii]
AID - 10.1016/j.cccn.2004.11.007 [doi]
PST - ppublish
SO  - Clin Chim Acta. 2005 Mar;353(1-2):193-9. doi: 10.1016/j.cccn.2004.11.007.