help button home button Biophys. J.
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Wenner, J. R.
Right arrow Articles by Bloomfield, V. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wenner, J. R.
Right arrow Articles by Bloomfield, V. A.

Biophys J, December 1999, p. 3234-3241, Vol. 77, No. 6

Crowding Effects on EcoRV Kinetics and Binding

Jay R. Wenner and Victor A. Bloomfield

Department of Biochemistry, University of Minnesota, St. Paul, Minnesota 55108 USA

The cytosol of the cell contains high concentrations of small and large macromolecules, but experimental data are often obtained in dilute solutions that do not reflect in vivo conditions. We have studied the crowding effect that large macromolecules have on EcoRV cleavage by adding high-molecular-weight Ficoll 70 to reaction solutions. Results indicate that Ficoll has surprisingly little effect on overall EcoRV reaction velocity because of offsetting increases in Vmax and Km, and stronger nonspecific binding. The changes in measured parameters can largely be attributed to the excluded volume effects on reactant activities and the slowing of protein diffusion. Covolume reduction upon binding appears to reinforce nonspecific binding strength, and kcat appears to be slowed by stronger nonspecific binding, which slows product release. The data also suggest that effective Ficoll particle volume decreases as its concentration increases above a few weight percent, which may be due to Ficoll interpenetration or compression.

Biophys J, December 1999, p. 3234-3241, Vol. 77, No. 6
© 1999 by the Biophysical Society   0006-3495/99/12/3234/08  $2.00


This article has been cited by other articles:


Home page
 Biophys. JHome page
G. Wieczorek and P. Zielenkiewicz
Influence of Macromolecular Crowding on Protein-Protein Association Rates--a Brownian Dynamics Study
Biophys. J., December 1, 2008; 95(11): 5030 - 5036.
[Abstract] [Full Text] [PDF]

Home page
 Biophys. JHome page
Z. Liu, W. Weng, R. M. Bookchin, V. L. Lew, and F. A. Ferrone
Free Energy of Sickle Hemoglobin Polymerization: A Scaled-Particle Treatment for Use with Dextran as a Crowding Agent
Biophys. J., May 1, 2008; 94(9): 3629 - 3634.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
A. Aranovich, G. Y. Gdalevsky, R. Cohen-Luria, I. Fishov, and A. H. Parola
Membrane-catalyzed Nucleotide Exchange on DnaA: EFFECT OF SURFACE MOLECULAR CROWDING
J. Biol. Chem., May 5, 2006; 281(18): 12526 - 12534.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Renal Physiol.Home page
D. Venturoli and B. Rippe
Ficoll and dextran vs. globular proteins as probes for testing glomerular permselectivity: effects of molecular size, shape, charge, and deformability
Am J Physiol Renal Physiol, April 1, 2005; 288(4): F605 - F613.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
B.-R. Zhou, Y. Liang, F. Du, Z. Zhou, and J. Chen
Mixed Macromolecular Crowding Accelerates the Oxidative Refolding of Reduced, Denatured Lysozyme: IMPLICATIONS FOR PROTEIN FOLDING IN INTRACELLULAR ENVIRONMENTS
J. Biol. Chem., December 31, 2004; 279(53): 55109 - 55116.
[Abstract] [Full Text] [PDF]

Home page
 Biophys. JHome page
M. Coppey, O. Benichou, R. Voituriez, and M. Moreau
Kinetics of Target Site Localization of a Protein on DNA: A Stochastic Approach
Biophys. J., September 1, 2004; 87(3): 1640 - 1649.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. M. Gonzalez, M. Jimenez, M. Velez, J. Mingorance, J. M. Andreu, M. Vicente, and G. Rivas
Essential Cell Division Protein FtsZ Assembles into One Monomer-thick Ribbons under Conditions Resembling the Crowded Intracellular Environment
J. Biol. Chem., September 26, 2003; 278(39): 37664 - 37671.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
A. H. Elcock
Atomic-level observation of macromolecular crowding effects: Escape of a protein from the GroEL cage
PNAS, March 4, 2003; 100(5): 2340 - 2344.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 1999 by the Biophysical Society.