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SILAC-based quantitative Nox4-Co-immunoprecipitation

ABSTRACT: Within the family of NADPH oxidases, Nox4 is unique as it is predominantly localized in the endoplasmic reticulum, has constitutive activity and generates H2O2. We hypothesize that these features are consequences of a so far unidentified Nox4-interacting protein. Interacting proteins were screened by quantitative SILAC-Co-immunoprecipitation in HEK293 cells stably overexpressing Nox4. By this technique, several interacting proteins were identified with calnexin showing the most robust interaction.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Ilka Wittig

LAB HEAD: Ralf Brandes

PROVIDER: PXD003514 | Pride | 2016-02-17

REPOSITORIES: Pride

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			Action	DRS
	140514_A1_KKP42_A2_01_FW.raw	Raw
	140514_A1_KKP42_A2_01_RW.raw	Raw
	140514_A1_KKP42_A2_02_FW.raw	Raw
	140514_A1_KKP42_A2_02_RW.raw	Raw
	140514_A1_KKP42_A2_03_FW.raw	Raw

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Publications

The Endoplasmic Reticulum Chaperone Calnexin Is a NADPH Oxidase NOX4 Interacting Protein.

Prior Kim-Kristin KK Wittig Ilka I Leisegang Matthias S MS Groenendyk Jody J Weissmann Norbert N Michalak Marek M Jansen-Dürr Pidder P Shah Ajay M AM Brandes Ralf P RP

The Journal of biological chemistry 20160209 13

Within the family of NADPH oxidases, NOX4 is unique as it is predominantly localized in the endoplasmic reticulum, has constitutive activity, and generates hydrogen peroxide (H2O2). We hypothesize that these features are consequences of a so far unidentified NOX4-interacting protein. Two-dimensional blue native (BN) electrophorese combined with SDS-PAGE yielded NOX4 to reside in macromolecular complexes. Interacting proteins were screened by quantitative SILAC (stable isotope labeling of amino a ...[more]

PMID: 26861875

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