Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Proc Natl Acad Sci U S A
2012 Jul 24;10930:12011-6. doi: 10.1073/pnas.1204129109.
Show Gene links
Show Anatomy links
A designed ankyrin repeat protein selected to bind to tubulin caps the microtubule plus end.
Pecqueur L
,
Duellberg C
,
Dreier B
,
Jiang Q
,
Wang C
,
Plückthun A
,
Surrey T
,
Gigant B
,
Knossow M
.
???displayArticle.abstract???
Microtubules are cytoskeleton filaments consisting of αβ-tubulin heterodimers. They switch between phases of growth and shrinkage. The underlying mechanism of this property, called dynamic instability, is not fully understood. Here, we identified a designed ankyrin repeat protein (DARPin) that interferes with microtubule assembly in a unique manner. The X-ray structure of its complex with GTP-tubulin shows that it binds to the β-tubulin surface exposed at microtubule (+) ends. The details of the structure provide insight into the role of GTP in microtubule polymerization and the conformational state of tubulin at the very microtubule end. They show in particular that GTP facilitates the tubulin structural switch that accompanies microtubule assembly but does not trigger it in unpolymerized tubulin. Total internal reflection fluorescence microscopy revealed that the DARPin specifically blocks growth at the microtubule (+) end by a selective end-capping mechanism, ultimately favoring microtubule disassembly from that end. DARPins promise to become designable tools for the dissection of microtubule dynamic properties selective for either of their two different ends.
Adams,
PHENIX: a comprehensive Python-based system for macromolecular structure solution.
2010, Pubmed
Adams,
PHENIX: a comprehensive Python-based system for macromolecular structure solution.
2010,
Pubmed
Bieling,
Reconstitution of a microtubule plus-end tracking system in vitro.
2007,
Pubmed
Bieling,
Fluorescence microscopy assays on chemically functionalized surfaces for quantitative imaging of microtubule, motor, and +TIP dynamics.
2010,
Pubmed
Bieling,
A minimal midzone protein module controls formation and length of antiparallel microtubule overlaps.
2010,
Pubmed
,
Xenbase
Binz,
Designing repeat proteins: well-expressed, soluble and stable proteins from combinatorial libraries of consensus ankyrin repeat proteins.
2003,
Pubmed
Binz,
High-affinity binders selected from designed ankyrin repeat protein libraries.
2004,
Pubmed
Bringmann,
A kinesin-like motor inhibits microtubule dynamic instability.
2004,
Pubmed
,
Xenbase
Brouhard,
XMAP215 is a processive microtubule polymerase.
2008,
Pubmed
,
Xenbase
Budde,
Analysis of microtubule polymerization in vitro and during the cell cycle in Xenopus egg extracts.
2006,
Pubmed
,
Xenbase
Buey,
The nucleotide switch of tubulin and microtubule assembly: a polymerization-driven structural change.
2006,
Pubmed
Desai,
Microtubule polymerization dynamics.
1997,
Pubmed
Dreier,
Rapid selection of high-affinity binders using ribosome display.
2012,
Pubmed
Dreier,
Her2-specific multivalent adapters confer designed tropism to adenovirus for gene targeting.
2011,
Pubmed
Du,
The kinesin-8 Kif18A dampens microtubule plus-end dynamics.
2010,
Pubmed
Emsley,
Coot: model-building tools for molecular graphics.
2004,
Pubmed
Fourniol,
Template-free 13-protofilament microtubule-MAP assembly visualized at 8 A resolution.
2010,
Pubmed
Goodwin,
Patronin regulates the microtubule network by protecting microtubule minus ends.
2010,
Pubmed
Gupta,
Plus end-specific depolymerase activity of Kip3, a kinesin-8 protein, explains its role in positioning the yeast mitotic spindle.
2006,
Pubmed
Hall,
Turbidity as a probe of tubulin polymerization kinetics: a theoretical and experimental re-examination.
2005,
Pubmed
Hanes,
In vitro selection and evolution of functional proteins by using ribosome display.
1997,
Pubmed
Helenius,
The depolymerizing kinesin MCAK uses lattice diffusion to rapidly target microtubule ends.
2006,
Pubmed
Honnappa,
Thermodynamics of the Op18/stathmin-tubulin interaction.
2003,
Pubmed
Hunter,
The kinesin-related protein MCAK is a microtubule depolymerase that forms an ATP-hydrolyzing complex at microtubule ends.
2003,
Pubmed
Knipling,
Preparation and properties of pure tubulin S.
1999,
Pubmed
Kollman,
Microtubule nucleation by γ-tubulin complexes.
2011,
Pubmed
Kramer,
Structural determinants for improved stability of designed ankyrin repeat proteins with a redesigned C-capping module.
2010,
Pubmed
Kueh,
Structural plasticity in actin and tubulin polymer dynamics.
2009,
Pubmed
Löwe,
Refined structure of alpha beta-tubulin at 3.5 A resolution.
2001,
Pubmed
Nawrotek,
The determinants that govern microtubule assembly from the atomic structure of GTP-tubulin.
2011,
Pubmed
Nogales,
Tubulin and FtsZ form a distinct family of GTPases.
1998,
Pubmed
Nogales,
High-resolution model of the microtubule.
1999,
Pubmed
Ravelli,
Insight into tubulin regulation from a complex with colchicine and a stathmin-like domain.
2004,
Pubmed
Rice,
The lattice as allosteric effector: structural studies of alphabeta- and gamma-tubulin clarify the role of GTP in microtubule assembly.
2008,
Pubmed
Scheel,
Purification and analysis of authentic CLIP-170 and recombinant fragments.
1999,
Pubmed
Silacci,
Gelsolin superfamily proteins: key regulators of cellular functions.
2004,
Pubmed
Steiner,
Efficient selection of DARPins with sub-nanomolar affinities using SRP phage display.
2008,
Pubmed
Varga,
Yeast kinesin-8 depolymerizes microtubules in a length-dependent manner.
2006,
Pubmed
Walczak,
XCTK2: a kinesin-related protein that promotes mitotic spindle assembly in Xenopus laevis egg extracts.
1997,
Pubmed
,
Xenbase
Walker,
Dynamic instability of individual microtubules analyzed by video light microscopy: rate constants and transition frequencies.
1988,
Pubmed
Wang,
Nucleotide-dependent bending flexibility of tubulin regulates microtubule assembly.
2005,
Pubmed
Wang,
Kif2C minimal functional domain has unusual nucleotide binding properties that are adapted to microtubule depolymerization.
2012,
Pubmed
Wear,
Capping protein: new insights into mechanism and regulation.
2004,
Pubmed
Weaver,
Kif18A uses a microtubule binding site in the tail for plus-end localization and spindle length regulation.
2011,
Pubmed
Yamashita,
Crystal structure of CapZ: structural basis for actin filament barbed end capping.
2003,
Pubmed
Zahnd,
Computational analysis of off-rate selection experiments to optimize affinity maturation by directed evolution.
2010,
Pubmed
