J. Biol. Chem., Vol. 263, Issue 33, 17429-17436, 11, 1988

Probing the mechanism of ATP hydrolysis on F-actin using vanadate and the structural analogs of phosphate BeF-3 and A1F-4

C Combeau and MF Carlier
Laboratoire d'Enzymologie, Centre National de la Recherche Scientifique, Gif-sur-Yvette, France.

The binding of phosphate analogs to F-ADP-actin filaments and their effect on the dynamics of the polymer have been investigated. Orthovanadate binds to F-actin with the same affinity as phosphate and, at low saturation levels, stabilizes the filament structure in a Pi- like fashion; at higher concentration, it promotes destabilization of the filament. BeF-3 and A1F-4 bind to F-ADP-actin in competition with Pi, with a stoichiometry of 1 mol/mol of F-actin subunit and an affinity 3 orders of magnitude higher than Pi (KD = 2 and 25 microM for BeF-3 and A1F-4, respectively). BeF-3 and A1F-4 mimic Pi in stabilizing F-actin and slow down the rate of actin dissociation from filaments 150- fold. Only 1% of F-ADP-BeF3 subunits provide extensive stabilization of the filament. A quantitative analysis of the stabilization by BeF-3 is proposed. While Pi appears in rapid equilibrium with F-ADP-actin, BeF-3 binds to and dissociates from F-ADP-actin at very slow rates (k+ = 4 M- 1 S-1; k = 8.10(-6) S-1). In addition, although functionally similar to the reconstituted F-ADP-Pi species, F-ADP-BeF3 has a different conformation as indicated by the 17% quenching of pyrenyl fluorescence linked to BeF-3 binding. We suggest that BeF-3 may be a good analog of the transition state F-ADP-P* and that Pi release following cleavage of ATP on F-actin might be rate-limited by the isomerization of F-ADP-P*.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				T. Inobe, K. Takahashi, K. Maki, S. Enoki, K. Kamagata, A. Kadooka, M. Arai, and K. Kuwajima Asymmetry of the GroEL-GroES Complex under Physiological Conditions as Revealed by Small-Angle X-Ray Scattering Biophys. J., February 15, 2008; 94(4): 1392 - 1402. [Abstract] [Full Text] [PDF]

				H. Liu, J. A. Enyeart, and J. J. Enyeart Angiotensin II inhibits native bTREK-1 K+ channels through a PLC-, kinase C-, and PIP2-independent pathway requiring ATP hydrolysis Am J Physiol Cell Physiol, August 1, 2007; 293(2): C682 - C695. [Abstract] [Full Text] [PDF]

				I. Fujiwara, D. Vavylonis, and T. D. Pollard Polymerization kinetics of ADP- and ADP-Pi-actin determined by fluorescence microscopy PNAS, May 22, 2007; 104(21): 8827 - 8832. [Abstract] [Full Text] [PDF]

				S. Romero, D. Didry, E. Larquet, N. Boisset, D. Pantaloni, and M.-F. Carlier How ATP Hydrolysis Controls Filament Assembly from Profilin-Actin: IMPLICATION FOR FORMIN PROCESSIVITY J. Biol. Chem., March 16, 2007; 282(11): 8435 - 8445. [Abstract] [Full Text] [PDF]

				A. Muhlrad, I. Ringel, D. Pavlov, Y. M. Peyser, and E. Reisler Antagonistic Effects of Cofilin, Beryllium Fluoride Complex, and Phalloidin on Subdomain 2 and Nucleotide-Binding Cleft in F-Actin Biophys. J., December 15, 2006; 91(12): 4490 - 4499. [Abstract] [Full Text] [PDF]

				E. C. Garner, C. S. Campbell, and R. D. Mullins Dynamic Instability in a DNA-Segregating Prokaryotic Actin Homolog Science, November 5, 2004; 306(5698): 1021 - 1025. [Abstract] [Full Text] [PDF]

				D. J. Crampton, S. Guo, D. E. Johnson, and C. C. Richardson The arginine finger of bacteriophage T7 gene 4 helicase: Role in energy coupling PNAS, March 30, 2004; 101(13): 4373 - 4378. [Abstract] [Full Text] [PDF]

				V. E. Galkin, A. Orlova, M. S. VanLoock, A. Shvetsov, E. Reisler, and E. H. Egelman ADF/cofilin use an intrinsic mode of F-actin instability to disrupt actin filaments J. Cell Biol., December 8, 2003; 163(5): 1057 - 1066. [Abstract] [Full Text] [PDF]

				P. Hilpela, P. Oberbanscheidt, P. Hahne, M. Hund, G. Kalhammer, J. V. Small, and M. Bahler SWAP-70 Identifies a Transitional Subset of Actin Filaments in Motile Cells Mol. Biol. Cell, August 1, 2003; 14(8): 3242 - 3253. [Abstract] [Full Text] [PDF]

				S. Wiesner, E. Helfer, D. Didry, G. Ducouret, F. Lafuma, M.-F. Carlier, and D. Pantaloni A biomimetic motility assay provides insight into the mechanism of actin-based motility J. Cell Biol., February 3, 2003; 160(3): 387 - 398. [Abstract] [Full Text] [PDF]

				U. K. Misra, G. Gawdi, and S. V. Pizzo Beryllium fluoride-induced cell proliferation: a process requiring P21ras-dependent activated signal transduction and NF-{kappa}B-dependent gene regulation J. Leukoc. Biol., March 1, 2002; 71(3): 487 - 494. [Abstract] [Full Text] [PDF]

				M. J. Dayel, E. A. Holleran, and R. D. Mullins Arp2/3 complex requires hydrolyzable ATP for nucleation of new actin filaments PNAS, December 18, 2001; 98(26): 14871 - 14876. [Abstract] [Full Text] [PDF]

				L. R. Otterbein, P. Graceffa, and R. Dominguez The Crystal Structure of Uncomplexed Actin in the ADP State Science, July 27, 2001; 293(5530): 708 - 711. [Abstract] [Full Text] [PDF]

				L. Xu and J. J. Enyeart Properties of ATP-dependent K+ channels in adrenocortical cells Am J Physiol Cell Physiol, January 1, 2001; 280(1): C199 - C215. [Abstract] [Full Text] [PDF]

				D. Cheng, J. Marner, and P. A. Rubenstein Interaction in Vivo and in Vitro between the Yeast Fimbrin, SAC6P, and a Polymerization-defective Yeast Actin (V266G and L267G) J. Biol. Chem., December 10, 1999; 274(50): 35873 - 35880. [Abstract] [Full Text] [PDF]

				L. Blanchoin and T. D. Pollard Mechanism of Interaction of Acanthamoeba Actophorin (ADF/Cofilin) with Actin Filaments J. Biol. Chem., May 28, 1999; 274(22): 15538 - 15546. [Abstract] [Full Text] [PDF]

				I. Gutsche-Perelroizen, J. Lepault, A. Ott, and M.-F. Carlier Filament Assembly from Profilin-Actin J. Biol. Chem., March 5, 1999; 274(10): 6234 - 6243. [Abstract] [Full Text] [PDF]

				L. D. Belmont, A. Orlova, D. G. Drubin, and E. H. Egelman A change in actin conformation associated with filament instability after Pi release PNAS, January 5, 1999; 96(1): 29 - 34. [Abstract] [Full Text] [PDF]

				V. Katanaev and M. Wymann GTPgammaS-induced actin polymerisation in vitro: ATP- and phosphoinositide-independent signalling via Rho-family proteins and a plasma membrane-associated guanine nucleotide exchange factor J. Cell Sci., January 6, 1998; 111(11): 1583 - 1594. [Abstract] [PDF]

				Y.-W. Xu, S. Morera, J. Janin, and J. Cherfils AlF3 mimics the transition state of protein phosphorylation in the crystal structure of nucleoside diphosphate kinase and MgADP PNAS, April 15, 1997; 94(8): 3579 - 3583. [Abstract] [Full Text] [PDF]

				B. Kuang and P. A. Rubenstein Beryllium Fluoride and Phalloidin Restore Polymerizability of a Mutant Yeast Actin (V266G,L267G) with Severely Decreased Hydrophobicity in a Subdomain 3/4 Loop J. Biol. Chem., January 10, 1997; 272(2): 1237 - 1247. [Abstract] [Full Text] [PDF]

				I. Perelroizen, D. Didry, H. Christensen, N.-H. Chua, and M.-F. Carlier Role of Nucleotide Exchange and Hydrolysis in the Function of Profilin in Actin Assembly J. Biol. Chem., May 24, 1996; 271(21): 12302 - 12309. [Abstract] [Full Text] [PDF]

				S. S. Rosenfeld, B. Rener, J. J. Correia, M. S. Mayo, and H. C. Cheung Equilibrium Studies of Kinesin-Nucleotide Intermediates J. Biol. Chem., April 19, 1996; 271(16): 9473 - 9482. [Abstract] [Full Text] [PDF]

				P. G. Allen, L. E. Laham, M. Way, and P. A. Janmey Binding of Phosphate, Aluminum Fluoride, or Beryllium Fluoride to F-actin Inhibits Severing by Gelsolin J. Biol. Chem., March 1, 1996; 271(9): 4665 - 4670. [Abstract] [Full Text] [PDF]

				D. Gopal and M. Burke Formation of Stable Inhibitory Complexes of Myosin Subfragment 1 Using Fluoroscandium Anions J. Biol. Chem., August 18, 1995; 270(33): 19282 - 19286. [Abstract] [Full Text] [PDF]

				T. Zhou, S. Radaev, B. P. Rosen, and D. L. Gatti Conformational Changes in Four Regions of the Escherichia coli ArsA ATPase Link ATP Hydrolysis to Ion Translocation J. Biol. Chem., August 3, 2001; 276(32): 30414 - 30422. [Abstract] [Full Text] [PDF]