一种用于测量来自尘螨嗜热栖尘螨的主要变应原Der p 1的半胱氨酸蛋白酶活性的灵敏荧光测定法。
A sensitive fluorescent assay for measuring the cysteine protease activity of Der p 1, a major allergen from the dust mite Dermatophagoides pteronyssinus.
作者信息
Schulz O, Sewell H F, Shakib F
机构信息
Division of Molecular and Clinical Immunology, Faculty of Medicine and Health Sciences, University of Nottingham, Queen's Medical Centre, Nottingham, UK.
出版信息
Mol Pathol. 1998 Aug;51(4):222-4. doi: 10.1136/mp.51.4.222.
Abstract
The potent allergenicity of Der p 1, a major allergen of the house dust mite Dermatophagoides pteronyssinus, is thought to be related to its cysteine protease activity. Therefore, there is considerable interest in developing a sensitive assay for measuring Der p 1 activity to screen for specific inhibitors. This study demonstrates for the first time that the activity of Der p 1 can be measured conveniently in a continuous rate assay with the fluorogenic substrate Boc-Gln-Ala-Arg-AMC (K(m) = 280 microM and kcat/K(m) = 4.6 x 10(3)/M/s).
摘要
屋尘螨嗜热栖尘螨的主要变应原Der p 1具有很强的变应原性，其被认为与半胱氨酸蛋白酶活性有关。因此，人们对开发一种灵敏的测定方法来检测Der p 1活性以筛选特异性抑制剂有着浓厚兴趣。本研究首次证明，使用荧光底物Boc-Gln-Ala-Arg-AMC（米氏常数K(m)=280微摩尔，催化常数与米氏常数的比值kcat/K(m)=4.6×10³/摩尔/秒），可以通过连续速率测定法方便地检测Der p 1的活性。
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