Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Science 9 May 2003:
Vol. 300. no. 5621, pp. 984 - 987
DOI: 10.1126/science.1083911
Prev | Table of Contents | Next

Reports

Recombinant Antibodies to the Small GTPase Rab6 as Conformation Sensors

Clément Nizak,1 Solange Monier,1 Elaine del Nery,1 Sandrine Moutel,2 Bruno Goud,1 Franck Perez1*

Here we report an approach, based on antibody phage display, to generate molecular conformation sensors. Recombinant antibodies specific to the guanosine triphosphate (GTP)–bound conformation of the small guanosine triphosphatase (GTPase) Rab6, a regulator of membrane traffic, were generated and used to locate Rab6·GTP in fixed cells, and, after green fluorescent protein (GFP) tagging and intracellular expression, to follow Rab6·GTP in vivo. Rab6 was in its GTP-bound conformation on the Golgi apparatus and transport intermediates, and the geometry of transport intermediates was modulated by Rab6 activity. More generally, the same approach could be applied to other molecules that can be locked in a particular conformation in vitro.

1 CNRS UMR144, Institut Curie, 26 rue d'Ulm, F75248 Paris Cedex 05, France.
2 Translational Department, Institut Curie, 26 rue d'Ulm, F75248 Paris Cedex 05, France.

* To whom correspondence should be addressed. E-mail: Franck.Perez{at}curie.fr

Read the Full Text


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Direct Selection of Monoclonal Phosphospecific Antibodies without Prior Phosphoamino Acid Mapping.
O. Vielemeyer, H. Yuan, S. Moutel, R. Saint-Fort, D. Tang, C. Nizak, B. Goud, Y. Wang, and F. Perez (2009)
J. Biol. Chem. 284, 20791-20795
   Abstract »    Full Text »    PDF »
mRNA Display Design of Fibronectin-based Intrabodies That Detect and Inhibit Severe Acute Respiratory Syndrome Coronavirus Nucleocapsid Protein.
H.-I. Liao, C. A. Olson, S. Hwang, H. Deng, E. Wong, R. S. Baric, R. W. Roberts, and R. Sun (2009)
J. Biol. Chem. 284, 17512-17520
   Abstract »    Full Text »    PDF »
Detection of GTP-Tubulin Conformation in Vivo Reveals a Role for GTP Remnants in Microtubule Rescues.
A. Dimitrov, M. Quesnoit, S. Moutel, I. Cantaloube, C. Pous, and F. Perez (2008)
Science 322, 1353-1356
   Abstract »    Full Text »    PDF »
Selection of Genetically Encoded Fluorescent Single Domain Antibodies Engineered for Efficient Expression in Escherichia coli.
A. Olichon and T. Surrey (2007)
J. Biol. Chem. 282, 36314-36320
   Abstract »    Full Text »    PDF »
Rab6 Regulates Both ZW10/RINT-1 and Conserved Oligomeric Golgi Complex-dependent Golgi Trafficking and Homeostasis.
Y. Sun, A. Shestakova, L. Hunt, S. Sehgal, V. Lupashin, and B. Storrie (2007)
Mol. Biol. Cell 18, 4129-4142
   Abstract »    Full Text »    PDF »
Expression of Ral GTPases, Their Effectors, and Activators in Human Bladder Cancer.
S. C. Smith, G. Oxford, A. S. Baras, C. Owens, D. Havaleshko, D. L. Brautigan, M. K. Safo, and D. Theodorescu (2007)
Clin. Cancer Res. 13, 3803-3813
   Abstract »    Full Text »    PDF »
The Rab4 effector Rabip4 plays a role in the endocytotic trafficking of Glut 4 in 3T3-L1 adipocytes.
M. Mari, P. Monzo, V. Kaddai, F. Keslair, T. Gonzalez, Y. Le Marchand-Brustel, and M. Cormont (2006)
J. Cell Sci. 119, 1297-1306
   Abstract »    Full Text »    PDF »
Cisternal Rab Proteins Regulate Golgi Apparatus Redistribution in Response to Hypotonic Stress.
S. Jiang and B. Storrie (2005)
Mol. Biol. Cell 16, 2586-2596
   Abstract »    Full Text »    PDF »
Regulation of Microtubule-dependent Recycling at the Trans-Golgi Network by Rab6A and Rab6A'.
J. Young, T. Stauber, E. del Nery, I. Vernos, R. Pepperkok, and T. Nilsson (2005)
Mol. Biol. Cell 16, 162-177
   Abstract »    Full Text »    PDF »
Recycling Endosomes Supply AMPA Receptors for LTP.
M. Park, E. C. Penick, J. G. Edwards, J. A. Kauer, and M. D. Ehlers (2004)
Science 305, 1972-1975
   Abstract »    Full Text »    PDF »
The C2 domains of the class I Rab11 family of interacting proteins target recycling vesicles to the plasma membrane.
A. J. Lindsay and M. W. McCaffrey (2004)
J. Cell Sci. 117, 4365-4375
   Abstract »    Full Text »    PDF »


To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)