A sensitive fluorescent assay for measuring the cysteine protease activity of Der p 1, a major allergen from the dust mite Dermatophagoides pteronyssinus

被引：45

作者：

Schulz, O ^{[1
]}

Sewell, HF ^{[1
]}

Shakib, F ^{[1
]}

机构：

[1] Univ Nottingham, Queens Med Ctr, Fac Med & Hlth Sci, Div Mol & Clin Immunol, Nottingham NG7 2UH, England

来源：

JOURNAL OF CLINICAL PATHOLOGY-MOLECULAR PATHOLOGY | 1998年 / 51卷 / 04期

关键词：

cysteine proteases; Der p 1; IgE;

D O I：

10.1136/mp.51.4.222

中图分类号：

R36 [病理学];

学科分类号：

100104 ;

摘要：

The potent allergenicity of Der p 1, a major allergen of the house dust Illite 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 mu M and k(cat)/K-m = 4.6 x 10(3)/M/s).

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收藏

页码：222 / 224

页数：3

相关论文

共 17 条

[1]

CHAMBERS I, 1997, BIOCHEM SOC T, V25, pS85

[2] SEQUENCE-ANALYSIS OF CDNA CODING FOR A MAJOR HOUSE DUST MITE ALLERGEN, DER-P-1 HOMOLOGY WITH CYSTEINE PROTEASES [J].

CHUA, KY ;

STEWART, GA ;

THOMAS, WR ;

SIMPSON, RJ ;

DILWORTH, RJ ;

PLOZZA, TM ;

TURNER, KJ .

JOURNAL OF EXPERIMENTAL MEDICINE, 1988, 167 (01) :175-182

[3] AUGMENTATION OF PERMEABILITY IN THE BRONCHIAL EPITHELIUM BY THE HOUSE-DUST MITE ALLERGEN DER P1 [J].

HERBERT, CA ;

KING, CM ;

RING, PC ;

HOLGATE, ST ;

STEWART, GA ;

THOMPSON, PJ ;

ROBINSON, C .

AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY, 1995, 12 (04) :369-378

[4] A MAJOR HOUSE-DUST MITE ALLERGEN DISRUPTS THE IMMUNOGLOBULIN-E NETWORK BY SELECTIVELY CLEAVING CD23 - INNATE PROTECTION BY ANTIPROTEASES [J].

HEWITT, CRA ;

BROWN, AP ;

HART, BJ ;

PRITCHARD, DI .

JOURNAL OF EXPERIMENTAL MEDICINE, 1995, 182 (05) :1537-1544

[5] CHARACTERIZATION OF THE PROTEASES IN THE CRUDE MITE EXTRACT [J].

INO, Y ;

ANDO, T ;

HAIDA, M ;

NAKAMURA, K ;

IWAKI, M ;

OKUDAIRA, H ;

MIYAMOTO, T .

INTERNATIONAL ARCHIVES OF ALLERGY AND APPLIED IMMUNOLOGY, 1989, 89 (04) :321-326

[6]

LOMBARDERO M, 1990, J IMMUNOL, V144, P1353

[7] Potential allergens stimulate the release of mediators of the allergic response from cells of mast cell lineage in the absence of sensitization with antigen-specific IgE [J].

Machado, DC ;

Horton, D ;

Harrop, R ;

Peachell, PT ;

Helm, BA .

EUROPEAN JOURNAL OF IMMUNOLOGY, 1996, 26 (12) :2972-2980

[8] On the potential significance of the enzymatic activity of mite allergens to immunogenicity. Clues to structure and function revealed by molecular characterization [J].

Robinson, C ;

Kalsheker, NA ;

Srinivasan, N ;

King, CM ;

Garrod, DR ;

Thompson, PJ ;

Stewart, GA .

CLINICAL AND EXPERIMENTAL ALLERGY, 1997, 27 (01) :10-21

[9] DER-P-I, A MAJOR ALLERGEN OF THE HOUSE-DUST MITE, PROTEOLYTICALLY CLEAVES THE LOW-AFFINITY RECEPTOR FOR HUMAN IGE (CD23) [J].

SCHULZ, O ;

LAING, P ;

SEWELL, HE ;

SHAKIB, F .

EUROPEAN JOURNAL OF IMMUNOLOGY, 1995, 25 (11) :3191-3194

[10] Cleavage of the low-affinity receptor for human IgE (CD23) by a mite cysteine protease: Nature of the cleaved fragment in relation to the structure and function of CD23 [J].

Schulz, O ;

Sutton, BJ ;

Beavil, RL ;

Shi, JG ;

Sewell, HF ;

Gould, HJ ;

Laing, P ;

Shakib, F .

EUROPEAN JOURNAL OF IMMUNOLOGY, 1997, 27 (03) :584-588