Cloning, characterization and expression of a calnexin homologue from the pathogenic fungus Paracoccidioides brasiliensis

Cloning, characterization and expression of a calnexin homologue from the pathogenic fungus Paracoccidioides brasiliensis

Author Feitosa, Luciano dos Santos Autor UNIFESP Google Scholar
Maria de Almeida Soares, Celia Google Scholar
Santos, Marcia Regina Machado Autor UNIFESP Google Scholar
Melo Bailao, Alexandre Google Scholar
Xander, Patricia Autor UNIFESP Google Scholar
Mortara, Renato Arruda Autor UNIFESP Google Scholar
Daniel Lopes, Jose Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Universidade Federal de Goiás (UFG)
Abstract We report the cloning of a Paracoccidioides brasiliensis cDNA, here named PbCnx, encoding the homologue of the endoplasmic reticulum molecular chaperone calnexin. Calnexin specifically recognizes monoglucosylated glycoproteins in the endoplasmic reticulum, thus being an essential component of the complex that interacts with the folded state of nascent secreted glycoproteins. the PbCnx open reading frame was found in a 1701 base pair (bp) fragment that encodes a 567 amino acid protein with an estimated mass of 62 680 Da. Northern and Southern blot hybridizations showed that PbCnx is encoded by a single, or a low number of, gene copies. PbCnx contains the hallmark KPEDWD motifs that are found in all members of the calnexin/calreticulin family proteins. A cDNA-encoding PbCnx was overexpressed as recombinant protein in Escherichia coli. the purified recombinant PbCnx was recognized by 6 out of 10 sera from PCM patients, a result that rules out its possible consideration for further use in diagnosis. Using confocal microscopy with anti-PbCnx mouse serum against yeast forms, a cytoplasmic staining pattern was observed. Copyright (c) 2006 John Wiley & Sons, Ltd.
Keywords chaperone
calnexin homologue
recombinant protein
paracoccidioidomycosis
Paracoccidioidides brasiliensis
Language English
Date 2007-02-01
Published in Yeast. Chichester: John Wiley & Sons Ltd, v. 24, n. 2, p. 79-87, 2007.
ISSN 0749-503X (Sherpa/Romeo, impact factor)
Publisher Wiley-Blackwell
Extent 79-87
Origin http://dx.doi.org/10.1002/yea.1438
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000244576600002
URI http://repositorio.unifesp.br/handle/11600/29469

Show full item record




File

File Size Format View

There are no files associated with this item.

This item appears in the following Collection(s)

Search


Browse

Statistics

My Account