Localization of Nox2 N-terminus using polyclonal antipeptide antibodies. - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Biochemical Journal Année : 2004

Localization of Nox2 N-terminus using polyclonal antipeptide antibodies.

Résumé

Nox2/gp91(phox) (where phox is phagocyte oxidase) is the catalytic membrane subunit of the granulocyte NADPH oxidase complex involved in host defence. The current model of membrane topology of Nox2 is based upon the identification of glycosylation sites, of regions that interact with the regulatory cytosolic factors and of the epitopes recognized by antibodies. So far, the localization of the N-terminus of Nox2 was only speculative. In order to clarify this localization, we raised a polyclonal antiserum against the N-terminal sequence M(1)GNWVAVNEGL(11). Purified antibodies recognize the mature protein as a broad band at 91 kDa (glycosylated form) or a band at 55 kDa after deglycosylation. Immunocytochemistry and flow-cytometry analysis show a strong binding of the anti-N-terminal antibodies to differentiated HL60 cells and neutrophils respectively, after permeabilization only. The N-terminus of Nox2 is therefore present in the mature protein and is located to the cytoplasmic side of the plasma membrane.

Domaines

Immunité innée

Dates et versions

hal-00820738 , version 1 (06-05-2013)

Identifiants

Citer

Marie-Hélène Paclet, Lydia M Henderson, Yannick Campion, Françoise Morel, Marie-Claire Dagher. Localization of Nox2 N-terminus using polyclonal antipeptide antibodies.. Biochemical Journal, 2004, 382 (Pt 3), pp.981-6. ⟨10.1042/BJ20040954⟩. ⟨hal-00820738⟩

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