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一种检测人肝脏丙氨酸:乙醛酸转氨酶的新型微量测定法。

A new micro-assay for human liver alanine: glyoxylate aminotransferase.

作者信息

Allsop J, Jennings P R, Danpure C J

机构信息

Division of Inherited Metabolic Diseases, Clinical Research Centre, Harrow, Middlesex, UK.

出版信息

Clin Chim Acta. 1987 Dec;170(2-3):187-93. doi: 10.1016/0009-8981(87)90127-6.

Abstract

A micro radiochemical method has been developed for the assay of the human liver peroxisomal enzyme alanine: glyoxylate aminotransferase (EC 2.6.1.44). The method, based on the electrophoretic separation of [14C]alanine (substrate) from [14C]pyruvate (product) is at least fifty times more sensitive than the currently-used spectrophotometric double enzyme method (Rowsell et al, Int J Biochem 1972;3: 247-257), enabling the enzymatic diagnosis of primary hyperoxaluria type 1 to be carried out on only 100 micrograms of human liver tissue obtained by percutaneous needle biopsy. The increased sensitivity of the new method allows the assay conditions to be such that they are on the linear parts of the time-course and protein concentration curves. This results in the activities of alanine: glyoxylate aminotransferase in human liver samples being 20-50% higher than those determined by the spectrophotometric method.

摘要

已开发出一种微量放射化学方法用于测定人肝脏过氧化物酶体酶丙氨酸

乙醛酸转氨酶(EC 2.6.1.44)。该方法基于[14C]丙氨酸(底物)与[14C]丙酮酸(产物)的电泳分离,其灵敏度比目前使用的分光光度双酶法(Rowsell等人,《国际生物化学杂志》1972年;3:247 - 257)至少高五十倍,使得仅通过经皮穿刺活检获得的100微克人肝脏组织就能进行1型原发性高草酸尿症的酶学诊断。新方法灵敏度的提高使得测定条件处于时间进程和蛋白质浓度曲线的线性部分。这导致人肝脏样品中丙氨酸:乙醛酸转氨酶的活性比用分光光度法测定的结果高20% - 50%。

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