Please use this identifier to cite or link to this item: http://repositorio.unifesp.br/handle/11600/38906
Title: Bradykinin Release Avoids High Molecular Weight Kininogen Endocytosis
Authors: Damasceno, Igor Z. [UNIFESP]
Melo, Kátia Regina Brasil [UNIFESP]
Nascimento, Fabio D.
Souza, Daianne S. P. [UNIFESP]
Araujo, Mariana S. [UNIFESP]
Souza, Sinval Estevam Gregorio [UNIFESP]
Sampaio, Misako Uemura [UNIFESP]
Nader, Helena Bonciani [UNIFESP]
Tersariol, Ivarne Luis dos Santos [UNIFESP]
Motta, Guacyara [UNIFESP]
Universidade Federal de São Paulo (UNIFESP)
Univ Anhanguera São Paulo UNIAN SP
Issue Date: 30-Mar-2015
Publisher: Public Library Science
Citation: Plos One. San Francisco: Public Library Science, v. 10, n. 3, 18 p., 2015.
Abstract: Human H-kininogen (120 kDa) plays a role in many pathophysiological processes and interacts with the cell surface through protein receptors and proteoglycans, which mediate H-kininogen endocytosis. in the present work we demonstrate that H-kininogen containing bradykinin domain is internalized and different endogenous kininogenases are present in CHO-K1 cells. We used CHO-K1 (wild type) and CHO-745 (mutant deficient in proteoglycans biosynthesis) cell lines. H-kininogen endocytosis was studied using confocal microscopy, and its hydrolysis by cell lysate fraction was determined by immunoblotting. Bradykinin release was also measured by radioimmunoassay. H-kininogen interaction with the cell surface of CHO-745 cells resulted in bradykinin release by serine proteases. in CHO-K1 cells, which produce heparan and chondroitin sulfate proteoglycans, internalization of H-kininogen through its bradykinin domain can occur on lipid raft domains/caveolae. Nevertheless bradykinin-free H-kininogen was not internalized by CHO-K1 cells. the H-kininogen present in acidic endosomal vesicles in CHO-K1 was approximately 10-fold higher than the levels in CHO-745. CHO-K1 lysate fractions were assayed at pH 5.5 and intact H-kininogen was totally hydrolyzed into a 62 kDa fragment. By contrast, at an assay pH 7.4, the remained fragments were 115 kDa, 83 kDa, 62 kDa and 48 kDa in size. the anti-pain-Sepharose chromatography separated endogenous kininogenases from CHO-K1 lysate fraction. No difference was detected in the assays at pH 5.5 or 7.4, but the proteins in the fraction bound to the resin released bradykinin from H-kininogen. However, the proteins in the unbound fraction cleaved intact H-kininogen at other sites but did not release bradykinin. H-kininogen can interact with extravascular cells, and is internalized dependent on its bradykinin domain and cell surface proteoglycans. After internalization, H-kininogen is proteolytically processed by intracellular kininogenases. the present data also demonstrates that serine or cysteine proteases in lipid raft domains/caveolae on the CHO cell can hydrolyze H-kininogen, thus releasing kinins.
URI: http://repositorio.unifesp.br/handle/11600/38906
ISSN: 1932-6203
Other Identifiers: http://dx.doi.org/10.1371/journal.pone.0121721
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Artigo

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