3-Hydroxy-3-methylglutaryl coenzyme A reductase activity in cultured fibroblasts from patients with mevalonate kinase deficiency: differential response to lipid supplied by fetal bovine serum in tissue culture medium.
3-Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase activity was measured in extracts of cultured fibroblasts derived from patients with mevalonate kinase deficiency (MKD). For six patients studied, the mean activity of 63.3 +/- 41.1 pmol/min-mg protein (+/- 1 SD, range 37.7-146.2) was significantly higher than the mean value in three control fibroblast lines of 11.1 +/- 3.5 (+/- 1 SD, range 8.0-14.9). These values were obtained using cells subcultured in medium supplemented with 10% fetal bovine serum (FBS) 21 h prior to assay. When cells were deprived of cholesterol by subculturing for 21 h in delipidated FBS, the mean value for patient cells was increased to 230.8 +/- 78.5 pmol/min-mg protein (range 130.9-333.8) as compared to 109.5 +/- 47.1 (range 78.0-163.6) for controls. The activity of HMG-CoA synthase in extracts of fibroblasts derived from the patients was not elevated. The mevalonic acid concentration in the surrounding culture medium was assessed by stable isotope dilution assay. For five patients, the mean concentration in medium containing FBS was 0.92 +/- 0.37 microM (+/- 1 SD, range 0.46-1.48) in contrast to 1.24 +/- 0.83 microM (range 0.46-2.54) for cells subcultured in delipidated FBS. The mean value for three control fibroblast lines was 0.22 +/- 0.12 microM (+/- 1 SD, range 0.11-0.35) for cells subcultured in FBS as compared to 0.01 +/- 0.01 microM (range 0.0-0.01 microM) for cells sucultured in delipidated FBS.(ABSTRACT TRUNCATED AT 250 WORDS)
Gibson
K M
KM
Kimberly H. Courtwright and Joseph W. Summers Metabolic Disease Center, Baylor University Medical Center, Dallas, TX 75246.
Hoffmann
G
G
Schwall
A
A
Broock
R L
RL
Aramaki
S
S
Sweetman
L
L
Nyhan
W L
WL
Brandt
I K
IK
Wappner
R S
RS
Lehnert
W
W
D000818:Animals; D001769:Blood; D002417:Cattle; D002478:Cells, Cultured; D002784:Cholesterol / Q000096:biosynthesis; D003470:Culture Media / Q000378:metabolism; D005347:Fibroblasts / Q000201:enzymology*; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D006903:Hydroxymethylglutaryl CoA Reductases / Q000378:metabolism*; D008055:Lipids / Q000494:pharmacology*; D008077:Lipoproteins, LDL / Q000378:metabolism; D008798:Mevalonic Acid / Q000378:metabolism; D010770:Phosphotransferases / Q000172:deficiency* / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor)
D003470:Culture Media; D008055:Lipids; D008077:Lipoproteins, LDL; D002784:Cholesterol; D006903:Hydroxymethylglutaryl CoA Reductases; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C021796:mevalonate kinase; D008798:Mevalonic Acid