Home | My Profile | Contact Us
Research Trends Products  |   order gateway  |   author gateway  |   editor gateway  
Register | Forgot Password

Author Resources
 Author Gateway
 Article submission guidelines

Editor Resources
 Editor/Referee Gateway

 Regional Subscription Agents/Distributors
Trends in Cell & Molecular Biology   Volumes    Volume 5 
Avoiding traps and pitfalls in enzyme inhibition studies: The crustacean gill (Na+, K+)-ATPase paradigm
Francisco A. Leone, Douglas C. Masui, Daniela P. Garçon, Carlos F. L. Fontes, John C. McNamara
Pages: 49 - 59
Number of pages: 11
Trends in Cell & Molecular Biology
Volume 5 

Copyright © 2010 Research Trends. All rights reserved

Acclimation of crustaceans to reduced salinities often results in newly-expressed ATPase activities other than (Na+, K+)-ATPase activity. In this study, we evaluate the inhibition of ATPase activity by vanadate, oligomycin, sodium azide, bafilomycin A1, ethacrynic acid, thapsigargin, EGTA, theophylline and levamisole in a microsomal fraction from the posterior gill tissue of the blue crab, Callinectes danae. Crabs were either acclimated for 10 days to a dilute medium of 15 ‰ salinity or were used fresh-caught (33‰ salinity) to standardize a protocol for estimating newly-expressed ATPase activities in the acclimated crabs. The (Na+, K+)-ATPase activity in the fresh-caught crab gills represents approximately 95% of the total gill ATPase activity. We thus used this preparation as a control to eliminate possible pitfalls and to compare the inhibition data obtained from the 15‰-acclimated crabs. By employing each different inhibitor alone or together with ouabain, our findings demonstrate the experimental protocol to constitute a reliable procedure by which to quantify the newly-expressed ATPase activities other than (Na+, K+)-ATPase that appear during the acclimation process. Our procedure is also well suited to avoid the putative non-specific inhibition that often arises during the steady-state kinetic characterization of crustacean gill (Na+, K+)-ATPases.
Buy this Article


Buy this article
Buy this volume
Subscribe to this title
Shopping Cart

Quick Links
Search Products
Browse in Alphabetical Order : Journals
Browse by Subject Classification : Journals

Ordering Information Ordering Information
Downloadable forms Downloadable Forms