alpha-chymotrypsin and Hypersensitivity

Extracts from ragweed pollen grains contain novel trypsin and chymotrypsin-like serine peptidases which are described in this report. The molecular mass of the chymotrypsin-like enzyme was 82 kDa, had a pH optimum near 9.0, and its activity was unaffected by chelating or reducing agents. It was inhibited by diisopropyl fluorophosphate (DFP), a general serine class inhibitor, and more specifically N-p-tosyl-L-phenylalanine chloromethyl ketone (TPCK), a chymotrypsin-like proteinase inhibitor. In addition to various synthetic substrates, the neuropeptides, vasoactive intestinal peptide (VIP) and substance P, which are required for normalized lung functions, were also rapidly hydrolysed. Activity toward protein substrates was not detected with the exception of the inactivation of alpha-1-proteinase inhibitor (alpha-1-PI) which occurred through cleavage within the reactive site loop. The 'trypsin-like' enzyme has a molecular mass near 80 kDa, a blocked N-terminus, a pH optimum near 9.0, and requires Ca++ for stability and activity, but not reducing agents. It is inhibited by DFP, and more specifically the trypsin-like proteinase inhibitor, N-p-tosyl-L-lysine chloromethyl ketone (TLCK). Again, activity toward protein substrates was not detected, but various synthetic substrates and biologically active peptides were efficiently cleaved. Significantly, atrial natriuretic peptide (ANP) and angiotensin 2 (ATII), whose degradation would amplify kinin activity and influence inflammatory diseases of the respiratory tract and nasal passages, were also rapidly hydrolyzed.

Topics: Allergens; Amino Acid Sequence; Asthma; Chymotrypsin; Humans; Hypersensitivity; Molecular Sequence Data; Molecular Weight; Peptides; Pollen; Serine Endopeptidases; Serine Proteinase Inhibitors; Substrate Specificity; Trypsin

One of the major allergenic proteins in the soybean 7S-globulin fraction, which was recognized by sera of about 25% soybean-sensitive patients with atopic dermatitis, was identified as alpha-subunit of beta-conglycinin. The IgE antibodies recognizing the alpha-subunit did not show a cross-reactivity against both alpha'- and beta-subunits known to be highly homologous to alpha-subunit, and also against other allergenic components identified in soybeans [Ogawa et al., J. Nutr. Sci. Vitaminol., 35, 555-565 (1993)]. The IgE-binding site(s) was estimated to be located on the peptide 232-283 of alpha-subunit.

Topics: Adolescent; Adult; Allergens; Antibodies, Anti-Idiotypic; Antigens, Plant; Child; Child, Preschool; Chymotrypsin; Cyanogen Bromide; Dermatitis, Atopic; Globulins; Glycine max; Humans; Hypersensitivity; Immunoglobulin E; Infant; Peptide Fragments; Plant Proteins; Seed Storage Proteins; Soybean Proteins

A new set of allergens from Dermatophagoides pteronyssinus and D. farinae (provisionally named DP5 and DF5, respectively) was isolated from the whole culture of mites. The apparent molecular weights of both allergens were shown to be 25,000 on SDS-PAGE under a reducing condition and 27,000 on Sephadex G-75 gel filtration chromatography. Both DP5 and DF5, as well as Der f III, possessed proteolytic activity. The results of substrate specificity and susceptibility to various protease inhibitors of DP5 and DF5 strongly suggested that they belonged to the chymotrypsin-like serine protease family. In sera from 88 mite-allergic patients, specific IgE antibodies to DP5 and/or DF5 were detected in only 41% of the sera by radio-allergosorbent test, while 90% and 93% had specific IgE antibodies to Der p I and/or Der f I and Der p II and/or Der f II, respectively.

Topics: Allergens; Amino Acid Sequence; Animals; Antigens, Dermatophagoides; Chymotrypsin; Glycoproteins; Humans; Hypersensitivity; Immunoglobulin E; Mites; Molecular Sequence Data; Radioallergosorbent Test; Substrate Specificity

A variety of dialyzable and non-dialyzable fractions from neutral timothy pollen extracts were studied in human and guinea pig RAST inhibition as well as in PCA. Extracted material fragmented by chemical reagents and by hydrolytic enzymes was also included in the study. It was found that some low molecular weight fractions were quite active in PCA and in RAST inhibition. The fragmented materials showed in a number of instances a considerably reduced inhibitory capacity in RAST but remarkably high PCA elicitation. Some of the enzymes did not produce significant modifications detectable by RAST inhibition although the test seems to be very sensitive to antigenic modification. It seems quite certain that a considerable number of antigenic specificities are involved in the clinical sera tested.

Topics: Allergens; Antigens; Chromatography, Affinity; Chromatography, Gel; Chymotrypsin; Dialysis; Hypersensitivity; Peptide Fragments; Peptide Hydrolases; Poaceae; Pollen; Radioimmunoassay; Trypsin

Topics: Adenosine Triphosphatases; Adenylyl Cyclases; Calcium; Cell Membrane; Cell Movement; Chymotrypsin; Cyclic AMP; Enzyme Precursors; Esterases; Histamine Release; Hypersensitivity; Mast Cells; Microtubules; Models, Biological; Phospholipases; Phospholipids; Prostaglandins

Topics: Antigens; Cell Migration Inhibition; Cell Movement; Centrifugation, Density Gradient; Chemotaxis; Chromatography, Gel; Chymotrypsin; Culture Media; Culture Techniques; Cytotoxicity Tests, Immunologic; Glycoproteins; Humans; Hypersensitivity; Lymphocytes; Molecular Weight; Monocytes; Neuraminidase

Topics: Adult; Allergens; Angioedema; Antigen-Antibody Reactions; Asthma; Autacoids; Chromatography, Gel; Chymotrypsin; Dermatitis, Atopic; Female; Histamine; Humans; Hypersensitivity; Immunization, Passive; Immunoglobulin E; Kinins; Male; Perfusion; Rhinitis, Allergic, Seasonal; Skin Tests; Sodium Chloride; Trypsin

Topics: Actinomyces; Animals; Antigen-Antibody Reactions; Antigens; Chymotrypsin; Culture Media; Esterases; Farmer's Lung; Humans; Hypersensitivity; Immune Sera; Immunoelectrophoresis; Oxidoreductases; Precipitins; Rabbits; Spores

Topics: Adult; Asthma; Chymotrypsin; Humans; Hypersensitivity; Male; Occupational Diseases; Powders; Respiratory Function Tests; Skin Tests; Trypsin

Topics: Aminocaproates; Animals; Antifibrinolytic Agents; Basophils; Chymotrypsin; Female; Fibrinolysis; Fibrinolytic Agents; Hypersensitivity; Male; Mast Cells; Microscopy, Electron; Nicotinic Acids; Pyridines; Pyridostigmine Bromide; Rabbits; Sodium Chloride; Streptodornase and Streptokinase; Vasodilator Agents

Topics: Ammonium Compounds; Anti-Bacterial Agents; Anti-Infective Agents, Local; Cataract; Cataract Extraction; Chymotrypsin; Cocaine; Cornea; Corneal Dystrophies, Hereditary; Corneal Ulcer; Drug Hypersensitivity; Drug Therapy; Edema; Epinephrine; Humans; Hypersensitivity; Keratitis; Keratoconjunctivitis; Mercury Compounds; Postoperative Complications; Quaternary Ammonium Compounds; Sutures; Tissue Adhesions; Toxicology

Topics: Anaphylaxis; Chymotrypsin; Hematologic Tests; Hypersensitivity; Obstetrics

Topics: Cataract Extraction; Chymotrypsin; Humans; Hypersensitivity

Topics: Anaphylaxis; Chymotrypsin; Hematologic Tests; Hypersensitivity; Immune System Diseases

Topics: Chymotrypsin; Hematologic Tests; Hypersensitivity; Medical Records; Trypsin

Topics: Anaphylaxis; Chymotrypsin; Hematologic Tests; Humans; Hypersensitivity

Topics: Chymotrypsin; Guinea Pigs; Histamine; Humans; Hypersensitivity; Immune System Diseases; Lung

Topics: Anaphylaxis; Chymotrypsin; Hematologic Tests; Hypersensitivity; Immune System Diseases; Water

Topics: Anaphylaxis; Chymotrypsin; Hematologic Tests; Hypersensitivity; Immune System Diseases; Medical Records

Topics: Chymotrypsin; Enzyme Therapy; Hematologic Tests; Hypersensitivity; Immune System Diseases; Trypsin

Topics: Chymotrypsin; Chymotrypsinogen; Enzyme Precursors; Hypersensitivity; Immune System Diseases; Trypsin; Trypsinogen

Topics: Chymotrypsin; Endopeptidases; Humans; Hydrolases; Hypersensitivity; Immune System Diseases; Peptide Hydrolases