铁螯合酶
色谱法
化学
高效液相色谱法
锌
孵化
超声
人口
生物化学
酶
血红素
医学
有机化学
环境卫生
作者
E. Rossi,K A Costin,P Garcia-Webb
出处
期刊:PubMed
日期:1988-12-01
卷期号:34 (12): 2481-5
被引量:38
摘要
We describe a new rapid, sensitive high-performance liquid-chromatographic (HPLC) method for determining ferrochelatase (EC 4.99.1.1) activity in lymphocytes. Zinc and mesoporphyrin are incubated aerobically with sonicated lymphocytes and the zinc mesoporphyrin formed is extracted with dimethyl sulfoxide-methanol-EDTA for quantification by HPLC. Incubation conditions, including the concentration of the palmitic acid activator, were optimized. The Michaelis constant (Km) was 2.1 mumol/L for mesoporphyrin, 22.2 mumol/L for zinc. The mean ferrochelatase activity (expressed as zinc mesoporphyrin formed per hour per milligram of lymphocyte protein) for a reference population was 3.25 (SD 0.43) nmol.h-1.mg-1. For three patients with erythrohepatic protoporphyria (EHP), activities were 1.11, 1.30, and 1.35. Neutrophils contain negligible ferrochelatase activity.
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