Affinity purification of alkylglycerol monooxygenase from rat liver microsomes by chimyl alcohol-Sepharose 4B column chromatography.
Alkylglycerol monooxygenase of rat liver microsomes was purified approximately to 97-fold with a 30% yield by procedures including affinity chromatography on chimyl alcohol-Sepharose 4B. Chimyl alcohol (1-O-hexadecylglycerol) was converted to the p-aminobenzylidene derivative and then coupled to 6-carboxyhexyl-Sepharose. The final enzyme preparation was in nearly a homogeneous state, judging from the results of sodium dodecyl sulfate-polyacrylamide disc gel electrophoresis, and it migrated to a position corresponding to an apparent molecular weight of 45,000. The results revealed that the native form of the enzyme (estimated to have a molecular weight of 400,000 as judged by Sepharose 6B column chromatography in a previous report, Ishibashi, T., and Y. Imai. 1983. Eur. J. Biochem. 132: 23-27) will polymerize to large aggregates.
D000818:Animals; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005990:Glycerol / Q000031:analogs & derivatives; D005995:Glyceryl Ethers; D008862:Microsomes, Liver / Q000201:enzymology*; D006899:Mixed Function Oxygenases / Q000302:isolation & purification*; D008970:Molecular Weight; D051381:Rats; D012685:Sepharose
D005995:Glyceryl Ethers; D012685:Sepharose; D006899:Mixed Function Oxygenases; C020678:glyceryl-ether monooxygenase; D005990:Glycerol; C004348:chimyl alcohol