title: Lytic water dynamics reveal evolutionarily conserved mechanisms of ATP hydrolysis by TIP49 AAA+ ATPases.
abstract: Eukaryotic TIP49a (Pontin) and TIP49b (Reptin) AAA+ ATPases play essential roles in key cellular processes. How their weak ATPase activity contributes to their important functions remains largely unknown and difficult to analyze because of the divergent properties of TIP49a and TIP49b proteins and of their homo- and hetero-oligomeric assemblies. To circumvent these complexities, we have analyzed the single ancient TIP49 ortholog found in the archaeon Methanopyrus kandleri (mkTIP49). All-atom homology modeling and molecular dynamics simulations validated by biochemical assays reveal highly conserved organizational principles and identify key residues for ATP hydrolysis. An unanticipated crosstalk between Walker B and Sensor I motifs impacts the dynamics of water molecules and highlights a critical role of trans-acting aspartates in the lytic water activation step that is essential for the associative mechanism of ATP hydrolysis.
authors.affiliation: Department of Biophysics, Saint Petersburg State Polytechnical University, Saint Petersburg 195251, Russia; Division of Molecular and Radiation Biophysics, Petersburg Nuclear Physics Institute, Gatchina 188300, Russia.
authors.forename: Arina
authors.initials: A
authors.lastname: Afanasyeva
authors.affiliation: Division of Biosciences, Institute for Structural and Molecular Biology, University College London, Darwin Building, Gower Street, London WC1E 6BT, UK.
authors.forename: Angela
authors.initials: A
authors.lastname: Hirtreiter
authors.affiliation: Division of Biosciences, Institute for Structural and Molecular Biology, University College London, Darwin Building, Gower Street, London WC1E 6BT, UK.
authors.forename: Anne
authors.initials: A
authors.lastname: Schreiber
authors.affiliation: Physikalische und Theoretische Chemie - NanoBioSciences, Technische Universität Braunschweig, Braunschweig 38106, Germany.
authors.forename: Dina
authors.initials: D
authors.lastname: Grohmann
authors.affiliation: Department of Biophysics, Saint Petersburg State Polytechnical University, Saint Petersburg 195251, Russia.
authors.forename: Georgii
authors.initials: G
authors.lastname: Pobegalov
authors.affiliation: Department of Chemistry, University College London, London WC1H 0AJ, UK.
authors.forename: Adam R
authors.initials: AR
authors.lastname: McKay
authors.affiliation: Division of Biosciences, Institute for Structural and Molecular Biology, University College London, Darwin Building, Gower Street, London WC1E 6BT, UK.
authors.forename: Irina
authors.initials: I
authors.lastname: Tsaneva
authors.affiliation: Department of Biophysics, Saint Petersburg State Polytechnical University, Saint Petersburg 195251, Russia; Division of Molecular and Radiation Biophysics, Petersburg Nuclear Physics Institute, Gatchina 188300, Russia.
authors.forename: Michael
authors.initials: M
authors.lastname: Petukhov
authors.affiliation: UMR 5099, CNRS, Toulouse F-31000, France; Laboratoire de Biologie Moléculaire Eucaryote, Université de Toulouse, Toulouse F-31000, France. Electronic address: kas@ibcg.biotoul.fr.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Käs
authors.affiliation: UMR 5099, CNRS, Toulouse F-31000, France; Laboratoire de Biologie Moléculaire Eucaryote, Université de Toulouse, Toulouse F-31000, France. Electronic address: grigor@biotoul.fr.
authors.forename: Mikhail
authors.initials: M
authors.lastname: Grigoriev
authors.affiliation: Division of Biosciences, Institute for Structural and Molecular Biology, University College London, Darwin Building, Gower Street, London WC1E 6BT, UK.
authors.forename: Finn
authors.initials: F
authors.lastname: Werner
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000235:genetics; D000255:Adenosine Triphosphate / Q000737:chemistry*; D019843:Archaeal Proteins / Q000737:chemistry* / Q000235:genetics; D001224:Aspartic Acid / Q000737:chemistry; D005075:Biological Evolution; D017124:Conserved Sequence; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D019605:Euryarchaeota / Q000737:chemistry* / Q000201:enzymology; D015870:Gene Expression; D006868:Hydrolysis; D056004:Molecular Dynamics Simulation; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D055503:Protein Multimerization; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D014867:Water / Q000737:chemistry*
chemical_list: D019843:Archaeal Proteins; D011994:Recombinant Proteins; D014867:Water; D001224:Aspartic Acid; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases