A yeast expression system for human galactose-1-phosphate uridylyltransferase.
نویسندگان
چکیده
منابع مشابه
Relationship between genotype, activity, and galactose sensitivity in yeast expressing patient alleles of human galactose-1-phosphate uridylyltransferase.
Impairment of the human enzyme galactose-1-phosphate uridylyltransferase (GALT) results in the potentially lethal disorder galactosemia; the biochemical basis of pathophysiology in galactosemia remains unknown. We have applied a yeast expression system for human GALT to test the hypothesis that genotype will correlate with GALT activity measured in vitro and with metabolite levels and galactose...
متن کاملEnzymatic micromethod for measuring galactose-1-phosphate uridylyltransferase activity in human erythrocytes.
A kinetic microspectrophotometric assay for galactose1-phosphate uridylyitransferase (EC 2.7.7. 12) activity in erythrocytes is described. BlOOd is collected in ammonium heparinized microhematocrit tubes, centrifuged and the erythrocytes are lysed with water. Galactose-1-phosphate uridylyltransferase activity is determined by mixing 25 ul of hemolysate with a reagent consisting of galactose1-ph...
متن کاملThe Q188R mutation in human galactose-1-phosphate uridylyltransferase acts as a partial dominant negative.
A longstanding goal in the fields of molecular genetics and biochemistry has been to explain how naturally occurring mutations associated with human metabolic disease impair activity of the enzymes involved. This goal is particularly complex for enzymes composed of multiple subunits, because single mutations may exert both intra- and intersubunit effects on holoenzyme structure and function. We...
متن کاملOrigins, distribution and expression of the Duarte-2 (D2) allele of galactose-1-phosphate uridylyltransferase
Duarte galactosemia is a mild to asymptomatic condition that results from partial impairment of galactose-1-phosphate uridylyltransferase (GALT). Patients with Duarte galactosemia demonstrate reduced GALT activity and carry one profoundly impaired GALT allele (G) along with a second, partially impaired GALT allele (Duarte-2, D2). Molecular studies reveal at least five sequence changes on D2 all...
متن کاملPurification of normal and inactive galactosemic galactose-1-phosphate uridylyltransferase from human red cells.
A rapid method is described for the purification of galactose-1-phosphate uridylyltransferase (EC 2.7.7.12) from human red blood cells by the use of DEAE-cellulose and two steps of affinity chromatography on a "uridine-aminohexyl" agarose column. The enzyme consists of two identical subunits of 31,000 molecular weight as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. C...
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ژورنال
عنوان ژورنال: Proceedings of the National Academy of Sciences
سال: 1993
ISSN: 0027-8424,1091-6490
DOI: 10.1073/pnas.90.2.398