Summary
Mevalonic aciduria due to mevalonate kinase deficiency, an inherited defect of cholesterol biosynthesis, has presented with clinical variability in 10 patients from 7 families. We sought to define a genetic basis for this heterogeneity by determining mevalonate kinase activity in fibroblast heterokaryons obtained by polyethylene glycol fusion. To this end we developed a DEAE-cellulose (Cl−) column chromatography procedure for assessing mevalonate kinase in cell extracts that would allow multiple rapid analyses. Fusion of control fibroblasts with those from affected patients from six families with mevalonate kinase deficiency yielded 37% of the mean control activity. None of the fusions between the six cell lines of patients resulted in measurable mevalonate kinase activity. Using the chromatographic procedure, we developed an optimized assay for mevalonate kinase in biopsied chorionic villi.K m values for chorionic villi were similar to those obtained in fibroblasts. Mevalonate kinase activity in biopsied chorionic villi showed a linear increase (0.75–4.3 nmol/min per mg protein) with gestational age from 7 to 14 weeks. Using the optimized assay in biopsied chorionic villi we performed a first-trimester prenatal diagnosis in a pregnancy at risk for mevalonate kinase deficiency and correctly diagnosed an unaffected fetus. The availability of an optimized assay for mevalonate kinase in biopsied chorionic villi should allow reliable first-trimester prenatal diagnosis for families at risk.
Similar content being viewed by others
References
Divry P, Rolland MO, Zabot MT, Mancini J, Philip N, Pinsard N (1991) Mevalonate kinase deficiency in 2 siblings.SSIEM, Proceedings of the 29th Annual Symposium, London, Sept. 1991, p 146.
Fukuda M., Tanaka A, Isshiki G (1990) Variation of lysosomal enzyme activity with gestational age in chorionic villi.J Inher Metab Dis 13: 862–866.
Gibson KM, Hoffmann G, Nyhan WL et al (1988) Mevalonate kinase deficiency in a child with cerebellar ataxia, hypotonia and mevalonic aciduria.Eur J Pediatr 148: 250–252.
Gibson KM, Lohr JL, Broock RL et al (1989) Mevalonate kinase in lysates of cultured human fibroblasts and lymphoblasts: Kinetic properties, assay conditions, carrier detection and measurement of residual activity in a patient with mevalonic aciduria.Enzyme 41: 47–55.
Gibson KM, Hoffmann G, Schwall A et al (1990) 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.J Lipid Res 31: 515–521.
Hoffmann G, Gibson KM, Brandt IK, Bader PI, Wappner RS, Sweetman L (1986) Mevalonic aciduria — an inborn error of cholesterol and nonsterol isoprene biosynthesis.N Engl J Med 314: 1610–1614
Hoffmann GF, Sweetman L, Bremer HJ et al (1991) Facts and artefacts in mevalonic aciduria: development of a stable isotope dilution GCMS assay for mevalonic acid and its application to physiological fluids, tissue samples, prenatal diagnosis and carrier detection.Clin Chim Acta 198: 209–228.
Kozich V, Gibson KM, Zeman J et al (1991) Mevalonic aciduria.J Inher Metab Dis 14: 265–266.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent.J Biol Chem 193: 265–275.
Page T, Bakay B, Nissinen E, Nyhan WL (1981) Hypoxanthine-guanine phosphoribosyltransferase variants: correlation of clinical phenotype with enzyme activity.J Inher Metab Dis 4: 203–206.
Peters JP, Wille W (1977) High yield mammalian cell fusion induced by polyethylene glycol. Pseudopodia are involved in the initiation of the fusion process.Cytobiology 15: 250–258.
Shin-Buehring YS, Osang M, Ziegler R, Schaub J (1976) A method for galactose-1-phosphate uridyltransferase assay and the separation of its isozymes by DEAE-cellulose column chromatography.Clin Chim Acta 70: 371–377.
Shin-Buehring YS, Beier T, Tan A, Osang M, Schaub J (1977a) The activity of galactose-1-phosphate uridyltransferase and galactokinase in human fetal organs.Pediatr Res 11: 1003–1009.
Shin-Buehring YS, Osang M, Ziegler R, Schaub J (1977b) A simple assay for galactokinase using DEAE-cellulose column chromatography.Clin Chim Acta 74: 1–5.
Sovik O, Sweetman L, Gibson KM, Nyhan WL (1984) Genetic complementation analysis of 3-hydroxy-3-methylglutaryl-coenzyme A lyase deficiency in cultured fibroblasts.Am J Hum Genet 36, 791–801.
Tanaka RD, Schafer BL, Lee LY, Freudenberger JS, Mosley ST (1990) Purification and regulation of mevalonate kinase from rat liver.J Biol Chem 265: 2391–2398.
Author information
Authors and Affiliations
Additional information
On leave from the Kimberly H. Courtwright and Joseph W. Summers Metabolic Disease Center and the Baylor Research Institute, Baylor University Medical Center, Dallas, TX, USA
Rights and permissions
About this article
Cite this article
Hoffmann, G.F., Brendel, S.U., Scharfschwerdt, S.R. et al. Mevalonate kinase assay using DEAE-cellulose column chromatography for first-trimester prenatal diagnosis and complementation analysis in mevalonic aciduria. J Inherit Metab Dis 15, 738–746 (1992). https://doi.org/10.1007/BF01800016
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01800016