Isolation of Cysteine Protease in the Crude Mite Extract, <i>Dermatophagoides farina</i><i>e</i>

Abstract

In order to study the relationship between cysteine protease and Der f l, which is one of the major allergens in the mite, Dermatophagoides farinae, isolation of cysteine protease was attempted using various column chromatographies. Both the potent cysteine protease activity and the allergenic activity were detected in the same fractions by anion exchange chromatography on a DEAE-Sephacel, gel chromatographies and chelating Sepharose 6B chromatography. In the double immunodiffusion test, the finally isolated fraction and rabbit anti-Der f I sera reacted to give a single precipitation line which fused completely with the precipitation line formed by Der f l and anti-Der f l sera. Sequence analysis for the first 10 N-terminal amino acids from cysteine protease and Der f l were identical. These results strongly suggest that cysteine protease of mites may be Der f l allergen and that measuring cysteine protease activity may possibly become a beneficial method for detecting Der f I allergens.

The role of protease activation of inflammation in allergic respiratory diseases

“…In 1989, Ino et al characterized Der p 1 as a cysteine protease, and this activity has been confirmed in Group 1 allergens from the other species of mites that cause allergy. [88][89][90][91] Group 3 mite allergens are serine proteases. [92][93][94][95][96] Additionally, mite extracts contain elastase, chymotrypsin-like protease, and other proteases.…”

Maturation of the activities of recombinant mite allergens Der p 1 and Der f 1, and its implication in the blockade of proteolytic activity

“…IgE‐binding activity for both group 1 and group 2 allergens is detected in the majority of sera from patients allergic to house dust mites. Group 1 allergens, which belong to the papain‐like cysteine protease family, are mainly found in mite feces and are thought to function as digestive enzymes in the intestine of mites [8–10]. IgE‐binding to the group 1 allergens is highly dependent on the tertiary structure of the allergens [5,11,12].…”

Crucial Commitment of Proteolytic Activity of a Purified Recombinant Major House Dust Mite Allergen Der p1 to Sensitization toward IgE and IgG Responses

“…Group 1 and group 2 allergens are major allergens derived from house dust mites based on the frequency of patients sensitized, amount of specific IgE, and content in mite extract (4 -10). The group 1 allergens, Der p 1 from D. pteronyssinus and Der f1 from D. farinae, belong to the papain-like cysteine protease family (11,12) and actually exhibit cysteine protease activity (13)(14)(15)(16). The cDNAs for Der p 1 and Der f 1 were isolated when cDNA cloning of allergens began (11,12,17).…”