期刊
ANALYTICAL BIOCHEMISTRY
卷 484, 期 -, 页码 102-104出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2015.05.019
关键词
Cell-free protein translation; Pancreatic microsomes; Mammalian ER membranes; Ribonuclease; Cyclotriazadisulfonamide (CADA); Translocation inhibitor
资金
- KU Leuven [GOA/10/014, PF/10/018]
- FWO [G.485.08]
Here, we demonstrate that pancreatic microsomal membranes from pigs, sheep, or cattle destined for human consumption can be used as a valuable and ethically correct alternative to dog microsomes for cell-free protein translocation. By adding adequate ribonuclease (RNase) inhibitors to the membrane fraction, successful in vitro co-translational translocation of wild-type and chimeric pre-prolactin into the lumen of rough microsomes was obtained. In addition, the human type I integral membrane proteins CD4 and VCAM-1 were efficiently glycosylated in RNase-treated microsomes. Thus, RNase-neutralized pancreatic membrane fractions from pig, cow, or sheep are a cheap, easily accessible, and fulfilling alternative to canine microsomes. (C) 2015 Elsevier Inc. All rights reserved.
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