Purification and some properties of rat intestinal ornithine decarboxylase.
Ornithine decarboxylase (ODC) was induced in rat small intestine by treatment with hypotonic solution in vitro and purified by two procedures, a conventional procedure and an immunoaffinity procedure. SDS-polyacrylamide gel electrophoresis showed that the molecular weight of the preparation purified by the immunoaffinity procedure (Mr = 53,000) was slightly larger than that of the preparation obtained by the conventional procedure (Mr = 52,000). Values for the Km for L-ornithine (0.1 mM), the isoelectric point (5.4), and the final specific activity (5.1-5.5 x 10(5) nmol CO2/mg protein/30 min) of the two preparations were similar to those reported for the rat liver ODC. Addition of a protease inhibitor (limabean trypsin inhibitor) to the crude extract prevented the appearance of the smaller enzyme (Mr = 52,000) obtained by the conventional purification procedure. Our result indicates that the large enzyme is native ODC and the smaller one is a partial proteolysis product of native ODC.
Miyamoto
K
K
Department of Nutrition, School of Medicine, Tokushima University.
Oka
T
T
Fujii
T
T
Yamaji
M
M
Minami
H
H
Nakabou
Y
Y
Hagihira
H
H
D000818:Animals; D005591:Chemical Fractionation; D055598:Chemical Phenomena; D002621:Chemistry; D002845:Chromatography / Q000379:methods; D000518:Eflornithine / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D007038:Hypotonic Solutions; D007124:Immunoenzyme Techniques; D007421:Intestine, Small / Q000201:enzymology*; D007526:Isoelectric Point; D008297:Male; D051379:Mice; D008970:Molecular Weight; D009955:Ornithine Decarboxylase / Q000302:isolation & purification*; D011480:Protease Inhibitors / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D014316:Tritium
D007038:Hypotonic Solutions; D011480:Protease Inhibitors; D014316:Tritium; D009955:Ornithine Decarboxylase; D000518:Eflornithine