title: Identification and analysis of occludin phosphosites: a combined mass spectrometry and bioinformatics approach.
abstract: The molecular function of occludin, an integral membrane component of tight junctions, remains unclear. VEGF-induced phosphorylation sites were mapped on occludin by combining MS data analysis with bioinformatics. In vivo phosphorylation of Ser490 was validated and protein interaction studies combined with crystal structure analysis suggest that Ser490 phosphorylation attenuates the interaction between occludin and ZO-1. This study demonstrates that combining MS data and bioinformatics can successfully identify novel phosphorylation sites from limiting samples.
authors.affiliation: Department of Cellular and Molecular Physiology, Penn State University College of Medicine, Hershey, Pennsylvania 17033, USA.
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Sundstrom
authors.forename: Brian R
authors.initials: BR
authors.lastname: Tash
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Murakami
authors.forename: John M
authors.initials: JM
authors.lastname: Flanagan
authors.forename: Maria C
authors.initials: MC
authors.lastname: Bewley
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Stanley
authors.forename: Kristin B
authors.initials: KB
authors.lastname: Gonsar
authors.forename: David A
authors.initials: DA
authors.lastname: Antonetti
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D019295:Computational Biology / Q000379:methods*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D062793:Occludin; D010750:Phosphoproteins / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D017433:Protein Structure, Secondary; D012694:Serine / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism; D062826:Zonula Occludens-1 Protein
chemical_list: D008565:Membrane Proteins; C568985:OCLN protein, human; D062793:Occludin; D010750:Phosphoproteins; C569042:TJP1 protein, human; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D062826:Zonula Occludens-1 Protein; D012694:Serine