H+-ATPase of crude homogenate of the outer mantle epithelium of Anodonta cygnea |
| |
Authors: | Oliveira P F Lopes I A Barrias C Rebelo da Costa A M |
| |
Institution: | Transporte Iónico e Fisiologia dos Gametas, CECA-UP, Porto, Portugal. |
| |
Abstract: | The outer mantle epithelium of the freshwater bivalve, Anodonta cygnea, is responsible for the mineralisation of the shell. Under short circuit conditions, it generates a current that is due to the operation of an electrogenic proton pump located in the apical barrier of that epithelium. Bafilomycin A1 and Concanamycin A inhibited the short circuit current. The IC50 and maximum inhibition dose were 0.17 and 0.5 microM for Bafilomycin A1, and 0.7 and 5 microM for Concanamycin A. The present work was undertaken to further characterise V-type ATPase of the outer mantle cells. The V-ATPase enzymatic activity of crude homogenate, measured as the amount of inorganic phosphorous released, due to ATP hydrolysis, in the presence of Na2SO3 (200 mM) was found to be 4.6+/-1.1 micromol Pi/mg protein/h, at 27 degrees C, pH 7.0-7.4 and ATP 4.5-6.0 microM. Bafilomycin A1 and Concanamycin A inhibit the V-ATPase activity with an IC50 of 14 and 8 nmol mg(-1), respectively. Dicyclohexylcarbodiimide (DCCD; 100 mM) and NaNO3 (100 microM) inhibited the V-type ATPase in what it seems a non-specific manner and NEM (100 mM) was unable to do it. Bafilomycin A1 (10 microM) and Concanamycin A (10 microM), inhibited 50-60% of the total activity. |
| |
Keywords: | Anodonta Freshwater bivalve V-ATPase Outer mantle epithelium Bafilomycin A1 Concanamycin A NEM DCCD |
本文献已被 ScienceDirect PubMed 等数据库收录! |
|