Cryoprotection of purified rat kidney transamidinase by polyethylene glycol.
Polyethylene glycol is a water-soluble polymer which is widely used in the pharmaceutical, cosmetic, and chemical industries. In this study, it is shown that polyethylene glycol is an effective cryoprotectant of rat kidney transamidinase purified from both the mitochondria and cytosol. Much of the activity is lost when the purified enzyme is frozen and thawed in sodium-potassium phosphate buffer in the absence of cryoprotectants. Polyethylene glycols with molecular weights of 4000 to 10,000 were effective cryoprotectants. However, polyethylene glycols with a molecular weight of 1000 or lower inhibited the purified enzyme. A concentration of only 0.01% polyethylene glycol 4000, 8000, or 10,000 was required for complete cryoprotection. In addition to polyethylene glycol, 0.5 mM ethylenediaminetetraacetic acid was required in the phosphate buffer for complete cryoprotection. The stabilization of purified transamidinase by polyethylene glycol will facilitate characterization experiments designed to compare the properties of the mitochondrial and cytosolic isozymes.
Tormanen
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Department of Chemistry, Central Michigan University, Mount Pleasant 48859.
D000579:Amidinotransferases / Q000302:isolation & purification*; D000818:Animals; D015925:Cryopreservation / Q000379:methods*; D003600:Cytosol / Q000201:enzymology; D005069:Evaluation Studies as Topic; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000302:isolation & purification; D007668:Kidney / Q000201:enzymology*; D008297:Male; D008928:Mitochondria / Q000201:enzymology; D008970:Molecular Weight; D011092:Polyethylene Glycols / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley
D007527:Isoenzymes; D011092:Polyethylene Glycols; D000579:Amidinotransferases; C020681:glycine amidinotransferase