title: Exploring the Protein Composition of the Plant Nuclear Envelope.
abstract: Due to rather limited sequence similarity, targeted identification of plant nuclear envelope and nuclear pore complex proteins has mainly followed two routes: (1) advanced computational identification followed by experimental verification and (2) immunoaffinity purification of complexes followed by mass spectrometry. Following candidate identification, fluorescence recovery after photobleaching (FRAP) and fluorescence resonance energy transfer (FRET) provide powerful tools to verify protein-protein interactions in situ at the NE. Here, we describe these methods for the example of Arabidopsis thaliana nuclear pore and nuclear envelope protein identification.
authors.affiliation: Department of Molecular Genetics, The Ohio State University, 520 Aronoff Laboratory, 318 West 12th Ave., Columbus, OH, 43210, USA.
authors.forename: Xiao
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Department of Botany, Kyoto University, Kyoto, Japan.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Tamura
authors.affiliation: Department of Biological and Medical Sciences, Oxford Brookes University, Oxford, UK.
authors.forename: Katja
authors.initials: K
authors.lastname: Graumann
authors.affiliation: Department of Molecular Genetics, The Ohio State University, 520 Aronoff Laboratory, 318 West 12th Ave., Columbus, OH, 43210, USA. meier.56@osu.edu.
authors.forename: Iris
authors.initials: I
authors.lastname: Meier
mesh_terms: D000818:Animals; D019295:Computational Biology / Q000379:methods; D036681:Fluorescence Recovery After Photobleaching; D031541:Fluorescence Resonance Energy Transfer; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D009685:Nuclear Envelope / Q000737:chemistry / Q000378:metabolism*; D028861:Nuclear Pore Complex Proteins / Q000235:genetics; D000071037:Place Cells / Q000378:metabolism*; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs / Q000187:drug effects; D025941:Protein Interaction Mapping; D060066:Protein Interaction Maps; D020543:Proteome; D040901:Proteomics / Q000379:methods; D017386:Sequence Homology, Amino Acid
chemical_list: D008565:Membrane Proteins; D028861:Nuclear Pore Complex Proteins; D020543:Proteome
keywords: Arabidopsis thaliana; Bioinformatics; Fluorescence recovery after photobleaching (FRAP); Fluorescence resonance energy transfer (FRET); Immunoaffinity purification; KASH protein; Nuclear pore complex