Quantitative, standardized assays for determining the concentrations of bovine lactoperoxidase, human salivary peroxidase, and human myeloperoxidase.
Quantitative, standardized assays for determining the concentrations of bovine lactoperoxidase, human salivary peroxidase, and human myeloperoxidase.
复制标题
用于测定牛乳过氧化物酶、人唾液过氧化物酶和人髓过氧化物酶浓度的定量标准化测定。
DOI：
10.1016/0003-2697(90)90220-4
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发表时间：
1990
影响因子：
2.9
通讯作者：
Rahemtulla,F
中科院分区：
文献类型：
--
作者：
Pruitt,KM;Kamau,DN;Miller,K;Månsson-Rahemtulla,B;Rahemtulla,F
由于过氧化物酶的重要生物学功能，人们对各种分泌物中过氧化物酶浓度的测量越来越感兴趣。目前还没有一种标准方法可用于对所报告的活动进行比较。本报告描述了可用于通过常用测定法测量过氧化物酶浓度的程序。已确定回归方程，可用于根据以下供体测定的活性计算牛乳过氧化物酶（LPO）、人唾液过氧化物酶（SPO）和人髓过氧化物酶（MPO）的浓度：连苯三酚、愈创木酚、2，2 ′-连氮双（3-乙基苄基噻唑啉-6-磺酸）和硫氰酸盐（SCN−）。这些供体的过氧化速率取决于单个测定中使用的过氧化氢（H2 O2）浓度，因此，为了获得准确、可重现的结果，必须仔细控制这些浓度。在ABTS、愈创木酚和连苯三酚试验中，通常存在于人唾液中的SCN-会通过简单的竞争动力学降低观察到的反应速率，而当在MPO的NBS试验中使用Cl-作为供体时，会增加观察到的速率。因此，在通过除硫代硝基苯甲酸（NBS）测定法以外的所有测定法测定过氧化物酶活性之前，必须从唾液样品中除去SCN−。LPO不能用作SPO或MPO的标准，因为LPO、SPO和MPO的比活性显著不同。
Because of the important biological functions of peroxidases, there is growing interest in the measurement of their concentrations in various secretions. At present, there is no standard method which allows for comparisons in reported activities. This report describes procedures which can be used to measure peroxidase enzyme concentrations by commonly employed assays. Regression equations have been determined which can be used to calculate concentrations of bovine lactoper-oxidase (LPO), human salivary peroxidase (SPO), and human myeloperoxidase (MPO) from activities measured with the following donors: pyrogallol, guaiacol, 2,2′-azinobis(3-ethylbenzylthiazoline-6-sulfonic acid), and thiocyanate (SCN−). The peroxidation rates of these donors depend upon the concentrations of hydrogen peroxide (H2O2) used in the individual assays and thus, for accurate, reproducible results, these concentrations must be carefully controlled. The SCN−normally present in human saliva will reduce observed reaction rates by simple competition kinetics in the ABTS, guaiacol and pyrogallol assays and will increase the rates observed when Cl−is used as a donor in NBS assay for MPO. Therefore, SCN−must be removed from saliva samples prior to peroxidase activity determination by all assays except the thionitrobenzoic acid (NBS) assay. LPO cannot be used as a standard for either SPO or MPO because the specific activities of LPO, SPO, and MPO are significantly different.