Purification of the microsomal Ca2+-ATPase from rat liver |
| |
Authors: | Yuh-Jiin Jong Adrian Sheldon Guo H Zhang Naomi Kraus-Friedmann |
| |
Affiliation: | (1) Department of Physiology and Cell Biology, University of Texas Medical School at Houston, 77225 Houston, Texas |
| |
Abstract: | Summary The Ca2+-ATPase from rat liver microsomes has been solubilized in Triton X-100 and purified to homogeneity by ficollsucrose treatment, column chromatography with agarose-hexane adenosine 5-triphosphate Type 2, and high pressure liquid chromatography (HPLC). The purified enzyme obtained by this sequential procedure exhibited a 183-fold increase in specific activity. After ficoll-sucrose treatment, the activity of the Ca2+-ATPase was stable for at least two weeks when stored at –70°C. In SDS-polyacrylamide gels, several fractions from HPLC chromatography showed a single band at a position corresponding to a molecular weight of about 107 kDa. This value is consistent with the molecular weight of the phosphoenzyme intermediate of endoplasmic reticulum (ER) Ca2+-ATPase. Further characterization of the ER Ca2+-ATPase was performed by western immunoblots. Antiserum raised against the 100-kDa sarcoplasmic reticulum (SR) Ca2+-ATPase cross-reacted with the purified Ca2+-ATPase from rat liver ER membranes. |
| |
Keywords: | Ca2+-ATPase endoplasmic reticulum rat liver purification |
本文献已被 SpringerLink 等数据库收录! |