title: Use of a Phosphatidylinositol Phosphate Affinity Chromatography (PIP Chromatography) for the Isolation of Proteins Involved in Protein Quality Control and Proteostasis Mechanisms in Plants.
abstract: Protein functionality depends directly on its accurately defined three-dimensional organization, correct and efficient posttranslational modification, and transport. However, proteins are continuously under a hostile environment threatening with folding aberrations, aggregation, and mistargeting. Therefore, proteins must be constantly "followed up" by a tightly regulated homeostatic mechanism specifically known as proteostasis. To this end other proteins ensure this close surveillance including chaperones as well as structural and functional members of the proteolytic mechanisms, mainly the autophagy and the proteasome related. They accomplish their action via interactions not only with other proteins but also with lipids as well as cytoskeletal components. We describe a protocol based on an affinity chromatographic approach aiming at the isolation of phosphatidyl inositol phosphate binding proteins, a procedure which results into the enrichment and purification of several members of the proteostasis mechanism, e.g. autophagy and proteasome, among other components of the cell signaling pathways.
authors.affiliation: CE.R.T.H.-IN.A.B., 6th km Charilaou-Thermi Rd., Thessaloniki, 570 01, Greece. mfarmaki@certh.gr.
authors.forename: T
authors.initials: T
authors.lastname: Farmaki
mesh_terms: D001343:Autophagy / Q000235:genetics; D002846:Chromatography, Affinity / Q000379:methods*; D008967:Molecular Biology / Q000379:methods*; D018129:Phosphatidylinositol Phosphates / Q000737:chemistry / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics / Q000302:isolation & purification*; D066329:Protein Aggregates / Q000235:genetics*; D017510:Protein Folding; D000074702:Proteostasis / Q000235:genetics
chemical_list: D018129:Phosphatidylinositol Phosphates; D010940:Plant Proteins; D066329:Protein Aggregates
keywords: Autophagy; Chaperones; Phosphatidylinositol phosphate (PIP); Proteasome; Sumoylation; Ubiquitin