title: Post translational modifications in adenovirus type 2.
abstract: We have combined 2-D SDS-PAGE with liquid chromatography-high resolving mass spectrometry (LC-MS) to explore the proteome of the adenovirus type 2 (Ad2) at the level of post translational modifications (PTMs). The experimental design included in-solution digestion, followed by titanium dioxide enrichment, as well as in-gel digestion of polypeptides after separation of Ad2 capsid proteins by 1-D and 2-D SDS-PAGE. All samples were analyzed using LC-MS with subsequent manual verification of PTM positions. The results revealed new phosphorylation sites that can explain the observed trains of protein spots observed for the pIII, pIIIa and pIV proteins. The pIIIa protein was found to be the most highly modified protein with now 18 verified sites of phosphorylation, three sites of nitrated tyrosine and one sulfated tyrosine. Nitrated tyrosines were also identified in pII. Lysine acetylations were detected in pII and pVI. The findings make the Ad2 virion much more complex than hitherto believed.
authors.affiliation: Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Rudbeck Laboratory, Uppsala University, SE-751 85 Uppsala, Sweden; Department of Chemistry-BMC, Science for Life Laboratory, Uppsala University, SE-751 24 Uppsala, Sweden. Electronic address: sara.lind@kemi.uu.se.
authors.forename: Sara
authors.initials: S
authors.lastname: Bergström Lind
authors.forename: Konstantin A
authors.initials: KA
authors.lastname: Artemenko
authors.forename: Lioudmila
authors.initials: L
authors.lastname: Elfineh
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Zhao
authors.forename: Jonas
authors.initials: J
authors.lastname: Bergquist
authors.forename: Ulf
authors.initials: U
authors.lastname: Pettersson
mesh_terms: D000107:Acetylation; D000256:Adenoviridae / Q000737:chemistry / Q000502:physiology*; D002853:Chromatography, Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D013058:Mass Spectrometry; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D020543:Proteome / Q000032:analysis; D014764:Viral Proteins / Q000378:metabolism*
chemical_list: D020543:Proteome; D014764:Viral Proteins
keywords: 2-D SDS-PAGE; Adenovirus type 2; High resolving mass spectrometry; Post translational modification analysis