一种基于一氧化碳生成量测定微粒体血红素加氧酶（EC 1.14.99.3）的新方法。

A new method for determination of microsomal haem oxygenase (EC 1.14.99.3) based on quantitation of carbon monoxide formation.

作者信息

Cavallin-Ståhl E, Jönsson G I, Lundh B

出版信息

Scand J Clin Lab Invest. 1978 Feb;38(1):69-76. doi: 10.3109/00365517809108405.

Abstract

Microsomal haem oxygenase (MHO) catalyses the main pathway for haem degradation. Hitherto bilirubin formation rate (BFR) has been used for determination of MHO activity. In the present study a method is described where MHO activity is assessed from the rate of carbon monoxide formation (VCO) in tissue homogenates with methaemalbumin as substrate. Formed CO is bound to haemoglobin present in the tissue homogenate. CO is measured by gas chromatography after reduction to methane. CO amounts of 0.01 nmol could be measured. This corresponded to an MHO activity of 0.05 pkat when using standard incubation mixture and 12 min incubation time. The correlation with MHO activity determined with the BFR technique was good (r = 0.94; n = 41); MHOVCO = 0.94-MHOBFR-0.08). Advantages with the VCO method are a ten-fold increase in sensitivity compared to the BFR method and independence of biliverdin reductase.

摘要

微粒体血红素加氧酶（MHO）催化血红素降解的主要途径。迄今为止，胆红素形成速率（BFR）一直用于测定MHO活性。在本研究中，描述了一种方法，即以高铁血红蛋白为底物，根据组织匀浆中一氧化碳形成速率（VCO）来评估MHO活性。生成的一氧化碳与组织匀浆中存在的血红蛋白结合。一氧化碳还原为甲烷后，通过气相色谱法进行测定。可测量的一氧化碳量为0.01 nmol。当使用标准孵育混合物和12分钟孵育时间时，这相当于0.05 pkat的MHO活性。与用BFR技术测定的MHO活性的相关性良好（r = 0.94；n = 41）；MHOVCO = 0.94 - MHOBFR - 0.08）。VCO方法的优点是与BFR方法相比灵敏度提高了十倍，并且不依赖于胆绿素还原酶。