A sensitive fluorescent assay for measuring the cysteine protease activity of Der p 1, a major allergen from the dust mite Dermatophagoides pteronyssinus.
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).
Schulz
O
O
Division of Molecular and Clinical Immunology, Faculty of Medicine and Health Sciences, University of Nottingham, Queen's Medical Centre, Nottingham, UK.
Sewell
H F
HF
Shakib
F
F
D000485:Allergens / Q000737:chemistry* / Q000302:isolation & purification; D000818:Animals; D039741:Antigens, Dermatophagoides; D003546:Cysteine Endopeptidases / Q000032:analysis*; D006023:Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification; D008925:Mites / Q000201:enzymology*; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity
D000485:Allergens; D039741:Antigens, Dermatophagoides; D006023:Glycoproteins; D003546:Cysteine Endopeptidases