The identification of an adenovirus receptor by using affinity capture and mass spectrometry

Citation:

Trauger SA, Wu E, Bark SJ, Nemerow GR, Siuzdak G. The identification of an adenovirus receptor by using affinity capture and mass spectrometry. ChembiochemChembiochemChembiochem. 2004;5 :1095-9.

Date Published:

Aug 6

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.

Notes:

Trauger, Sunia AWu, EugeneBark, Steve JNemerow, Glen RSiuzdak, GaryEY11431/EY/NEI NIH HHS/R01 GM 5-71094/GM/NIGMS NIH HHS/GermanyChembiochem. 2004 Aug 6;5(8):1095-9.