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The Identification of an Adenovirus Receptor by Using Affinity Capture and Mass Spectrometry
Author(s) -
Trauger Sunia A.,
Wu Eugene,
Bark Steve J.,
Nemerow Glen R.,
Siuzdak Gary
Publication year - 2004
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200400037
Subject(s) - receptor , glycoprotein , western blot , tandem mass spectrometry , blot , affinity chromatography , chemistry , mass spectrometry , membrane protein , microbiology and biotechnology , biochemistry , biology , chromatography , gene , membrane , enzyme
Abstract A tandem mass spectrometry‐based approach is demonstrated for detecting a receptor for Ad37, one of the causative agents for epidemic keratoconjunctivitis. Partial purification of membrane glycoproteins was performed by using lectin‐affinity chromatography and SDS‐PAGE. Gel bands that were shown to bind Ad37 by using Viral Overlay Protein Blot Assay (VOPBA) were excised, proteolyzed and analyzed by using nanoLC‐MS/MS to identify putative receptors contained in a mixture of proteins. Four candidate receptors were identified among approximately 50 proteins based on a search against a protein database. Inhibition of gene delivery mediated by an Ad37 vector, with antibodies against the glycoproteins identified by tandem mass spectrometry, strongly indicated that Membrane Cofactor Protein (MCP), a member of the complement regulatory family of proteins, is the receptor. This rapid and sensitive MS/MS‐based strategy is perceived to have wide potential applications for the detection of viral receptors.