Mevalonate kinase assay using DEAE‐cellulose column chromatography for first‐trimester prenatal diagnosis and complementation analysis in mevalonic aciduria

Autor: Georg F. Hoffmann, S. R. Scharfschwerdt, Yoon S. Shin, K. M. Gibson, S. U. Brendel, I. M. Speidel
Rok vydání: 1991
Předmět:
medicine.medical_specialty
Mevalonic Acid
Prenatal diagnosis
Chromatography
DEAE-Cellulose
03 medical and health sciences
0302 clinical medicine
Pregnancy
Prenatal Diagnosis
Internal medicine
Genetics
medicine
Humans
Fibroblast
reproductive and urinary physiology
Genetics (clinical)
030304 developmental biology
0303 health sciences
Fetus
Mevalonate kinase deficiency
biology
Phosphotransferases
Mevalonate kinase
Metabolism
Fibroblasts
Chromatography
Ion Exchange
medicine.disease
Molecular biology
3. Good health
Phosphotransferases (Alcohol Group Acceptor)
Pregnancy Trimester
First
medicine.anatomical_structure
Endocrinology
Mevalonic aciduria
embryonic structures
biology.protein
Chorionic villi
Female
Chromatography
Thin Layer
Chorionic Villi
Metabolism
Inborn Errors
030217 neurology & neurosurgery
Zdroj: Journal of Inherited Metabolic Disease. 15:738-746
ISSN: 1573-2665
0141-8955
DOI: 10.1007/bf01800016
Popis: 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. Km 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.
Databáze: OpenAIRE
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