title: Structure and flexibility of the endosomal Vps34 complex reveals the basis of its function on membranes.
abstract: Phosphatidylinositol 3-kinase Vps34 complexes regulate intracellular membrane trafficking in endocytic sorting, cytokinesis, and autophagy. We present the 4.4 angstrom crystal structure of the 385-kilodalton endosomal complex II (PIK3C3-CII), consisting of Vps34, Vps15 (p150), Vps30/Atg6 (Beclin 1), and Vps38 (UVRAG). The subunits form a Y-shaped complex, centered on the Vps34 C2 domain. Vps34 and Vps15 intertwine in one arm, where the Vps15 kinase domain engages the Vps34 activation loop to regulate its activity. Vps30 and Vps38 form the other arm that brackets the Vps15/Vps34 heterodimer, suggesting a path for complex assembly. We used hydrogen-deuterium exchange mass spectrometry (HDX-MS) to reveal conformational changes accompanying membrane binding and identify a Vps30 loop that is critical for the ability of complex II to phosphorylate giant liposomes on which complex I is inactive.
authors.affiliation: MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.
authors.forename: Ksenia
authors.initials: K
authors.lastname: Rostislavleva
authors.affiliation: MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Soler
authors.affiliation: MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.
authors.forename: Yohei
authors.initials: Y
authors.lastname: Ohashi
authors.affiliation: MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.
authors.forename: Lufei
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Structural Biology Research Center, VIB, B-1050 Brussels, Belgium.|Structural Biology Brussels, Vrije Universiteit Brussel, B-1050 Brussel, Belgium.
authors.forename: Els
authors.initials: E
authors.lastname: Pardon
authors.affiliation: MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.
authors.forename: John E
authors.initials: JE
authors.lastname: Burke
authors.affiliation: MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.
authors.forename: Glenn R
authors.initials: GR
authors.lastname: Masson
authors.affiliation: MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.
authors.forename: Chris
authors.initials: C
authors.lastname: Johnson
authors.affiliation: Structural Biology Research Center, VIB, B-1050 Brussels, Belgium.|Structural Biology Brussels, Vrije Universiteit Brussel, B-1050 Brussel, Belgium.
authors.forename: Jan
authors.initials: J
authors.lastname: Steyaert
authors.affiliation: The Babraham Institute, Cambridge UK.
authors.forename: Nicholas T
authors.initials: NT
authors.lastname: Ktistakis
authors.affiliation: MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.
authors.forename: Roger L
authors.initials: RL
authors.lastname: Williams
mesh_terms: D002462:Cell Membrane / Q000737:chemistry / Q000201:enzymology*; D058546:Class III Phosphatidylinositol 3-Kinases / Q000737:chemistry / Q000648:ultrastructure*; D018360:Crystallography, X-Ray; D011992:Endosomes / Q000737:chemistry / Q000201:enzymology*; D055503:Protein Multimerization; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D058571:Vacuolar Sorting Protein VPS15 / Q000737:chemistry / Q000648:ultrastructure
chemical_list: D058546:Class III Phosphatidylinositol 3-Kinases; D058571:Vacuolar Sorting Protein VPS15