Purification and some properties of an oxydative inhibitor in rabbit reticulocyte lysates.
Protein synthesis in rabbit reticulocyte lysates in the presence of heme is inhibited by 50% by the addition of 4 mM GSSG (oxidized glutathione). The incubation of the rabbit reticulocyte lysate with 4 mM GSSG at 30 degrees C for 30 min will cause activation of an inhibitor of protein synthesis which could be purified from the lysates through a five-step procedure. The inhibitor results in a 70-80% inhibition after a 1 h incubation. The inhibitor consists of one polypeptide of 23 kDa apparent molecular weight and is 90% pure as judged by sodium dodecyl sulfate/polyacrylamide gel electrophoresis. However, in the presence of cAMP (10 mM) or GEF (guanine nucleotide exchange factor) (0.3 microgram), protein synthesis in the inhibited reticulocyte lysate will be already recovered.
Erdogdu
G
G
Higher Educational Council of Turkey, Ankara, Turkey. egonul@rorqual.cc.metu.edu.tr
Dholakia
J N
JN
Wahba
A J
AJ
D000818:Animals; D001798:Blood Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002474:Cell-Free System; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D000242:Cyclic AMP / Q000494:pharmacology; D019803:Glutathione Disulfide / Q000494:pharmacology*; D020662:Guanine Nucleotide Exchange Factors; D007700:Kinetics; D010084:Oxidation-Reduction; D011500:Protein Synthesis Inhibitors / Q000097:blood* / Q000302:isolation & purification; D011506:Proteins / Q000378:metabolism; D011817:Rabbits; D012156:Reticulocytes / Q000187:drug effects / Q000378:metabolism*
D001798:Blood Proteins; D020662:Guanine Nucleotide Exchange Factors; D011500:Protein Synthesis Inhibitors; D011506:Proteins; D000242:Cyclic AMP; D019803:Glutathione Disulfide