title: A chemical glycoproteomics platform reveals O-GlcNAcylation of mitochondrial voltage-dependent anion channel 2.
abstract: Protein modification by O-linked β-N-acetylglucosamine (O-GlcNAc) is a critical cell signaling modality, but identifying signal-specific O-GlcNAcylation events remains a significant experimental challenge. Here, we describe a method for visualizing and analyzing organelle- and stimulus-specific O-GlcNAcylated proteins and use it to identify the mitochondrial voltage-dependent anion channel 2 (VDAC2) as an O-GlcNAc substrate. VDAC2(-/-) cells resist the mitochondrial dysfunction and apoptosis caused by global O-GlcNAc perturbation, demonstrating a functional connection between O-GlcNAc signaling and mitochondrial physiology through VDAC2. More broadly, our method will enable the discovery of signal-specific O-GlcNAcylation events in a wide array of experimental contexts.
authors.affiliation: Department of Chemistry, University of California, Berkeley, Berkeley, CA 94720, USA.
authors.forename: Krishnan K
authors.initials: KK
authors.lastname: Palaniappan
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Hangauer
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Smith
authors.forename: Brian P
authors.initials: BP
authors.lastname: Smart
authors.forename: Austin A
authors.initials: AA
authors.lastname: Pitcher
authors.forename: Emily H
authors.initials: EH
authors.lastname: Cheng
authors.forename: Carolyn R
authors.initials: CR
authors.lastname: Bertozzi
authors.forename: Michael
authors.initials: M
authors.lastname: Boyce
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D016521:Electrophoresis, Gel, Pulsed-Field; D005456:Fluorescent Dyes / Q000737:chemistry; D006023:Glycoproteins / Q000378:metabolism; D006031:Glycosylation; D057809:HEK293 Cells; D006801:Humans; D019169:Jurkat Cells; D051379:Mice; D008928:Mitochondria / Q000378:metabolism*; D040901:Proteomics; D013379:Substrate Specificity; D050996:Voltage-Dependent Anion Channel 2 / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*
chemical_list: D005456:Fluorescent Dyes; D006023:Glycoproteins; C494765:VDAC2 protein, human; D050996:Voltage-Dependent Anion Channel 2; D000117:Acetylglucosamine