Aminopeptidase activity in human nasal mucosa

Autor: Kimihiro Ohkubo, Marco Merida, Louis B. Hersh, James N. Baraniuk, Michael A. Kaliner, Robert J. Hohman
Rok vydání: 1998
Předmět:
Cytosol alanyl aminopeptidase
medicine.medical_specialty
Immunology
Mucous membrane of nose
NEP:
Neutral endopeptidase
CD13 Antigens
Biology
Turbinates
Aminopeptidases
Binding
Competitive
Aminopeptidase
Article
ACE:
Angiotensin-converting enzyme
Substrate Specificity
Puromycin-Sensitive Aminopeptidase
Leucyl Aminopeptidase
rhinitis
Leucine
Ala-NA:
Alanine-p -nitroanilide
Internal medicine
Enkephalin-degrading aminopeptidase
medicine
Humans
Immunology and Allergy
Protease Inhibitors
Zymography
puromycin sensitive aminopeptidase
ApM:
Aminopeptidase M
Antigens
Leucyl aminopeptidase
aminopeptidase M
Histocytochemistry
Ala-MNA:
l -alanine-4-methoxy-2-naphthylamide
Leu-Ap:
Leucine-enkephalin–degrading aminopeptidase
Nasal Lavage Fluid
Immunohistochemistry
PS-Ap:
Puromycin-sensitive aminopeptidase
Nasal Mucosa
Endocrinology
glandular secretion
Electrophoresis
Polyacrylamide Gel
Puromycin
Alanine aminopeptidase
Ala-Ap:
Alanine aminopeptidase
bradykinin
Subcellular Fractions
Zdroj: The Journal of Allergy and Clinical Immunology
ISSN: 0091-6749
DOI: 10.1016/s0091-6749(98)70013-2
Popis: Background: Aminopeptidases activate bradykinin and degrade many inflammatory peptides. Objective: The objective of this study was to identify the types of aminopeptidase activities in human nasal mucosa. Methods: Human nasal mucosa was homogenized (n = 12), and cytoplasmic (S2) and membrane-rich (P2) fractions were obtained. Several aminopeptidase (Ap) activities were defined by (1) substrate specificity with leucine-enkephalin (leu-Ap) and alanine-nitroanilide (ala-Ap), (2) inhibitor studies with puromycin and bestatin, (3) enzyme activity histochemistry (zymography), (4) immunohistochemistry, and (5) gel electrophoresis. Human volunteers had methacholine, histamine, and allergen nasal provocations to determine the mechanisms controlling nasal aminopeptidase secretion in vivo. Results: P2 was the largest reservoir of puromycin-resistant aminopeptidase activity (630 pmol leu-enk/min/mg protein). S2 contained 32 pmol leu-enk/min/mg activity, with 80% representing puromycin-resistant activity and 20% puromycin-sensitive aminopeptidase (PS-Ap). Ala-Ap was detected in both P2 and S2 fractions and was localized by zymography to epithelial and gland cells. Anti–rat brain–soluble PS-Ap IgG detected immunoreactive material in epithelium, glands, and endothelium. In nasal provocation studies, leu-AP correlated with glandular exocytosis but not vascular leak. Conclusions: The predominant aminopeptidase in human nasal epithelial and submucosal gland cells was membrane-bound puromycin-resistant aminopeptidase. A novel soluble puromycin-resistant aminopeptidase and lower amounts of soluble PS-Ap were also detected. (J Allergy Clin Immunol 1998;102:741-50.)
Databáze: OpenAIRE
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