Ethanolamine base-exchange reaction in rat brain microsomal subfractions.
Crude microsomal fractions have been subfractionated by differential ultracentrifugation into subfractions A, B, and C, corresponding to light smooth, heavy smooth, and rough microsomal membranes, respectively. The purity and the vesiculation of the membranes were checked biochemically. Subfraction C showed the highest ethanolamine base-exchange activity, both on phospholipid and protein bases. The other two subfractions had roughly similar activities. The kinetic behavior of the enzyme activity, although anomalous, was similar in the three subfractions. Treatment of the vesicles with Pronase or with mercury-dextran produced inactivation of the ethanolamine base-exchange reaction in the three subfractions. These findings suggest that the active site of base-exchange activity would be localized on the external leaflet of the vesicles. Treatment of the membranes with trinitrobenzenesulfonic acid (TNBS) has shown that the newly synthesized phosphatidylethanolamine (PE) belongs to a pool easily reacting with the probe, independent of the subfraction investigated. On the other hand, the distribution of the bulk membrane PE reacting with TNBS differs in the three subfractions examined. It is concluded that the newly synthesized PE and probably the active site of the enzyme are on the external leaflet of the membrane in all subfractions and that the ethanolamine base-exchange reaction has similar properties in all subfractions.
L
L
Corazzi
G
G
Porcellati
L
L
Freysz
L
L
Binaglia
R
R
Roberti
G
G
Arienti
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002784:Cholesterol / Q000032:analysis; D003911:Dextrans / Q000494:pharmacology; D003576:Electron Transport Complex IV / Q000378:metabolism; D019856:Ethanolamine; D004983:Ethanolamines / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D008628:Mercury / Q000494:pharmacology; D008861:Microsomes / Q000032:analysis / Q000187:drug effects / Q000378:metabolism*; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism; D009941:Organomercury Compounds; D010714:Phosphatidylethanolamines / Q000378:metabolism; D010743:Phospholipids / Q000032:analysis; D011402:Pronase / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism; D014302:Trinitrobenzenesulfonic Acid / Q000494:pharmacology
D003911:Dextrans; D004983:Ethanolamines; D009941:Organomercury Compounds; D010714:Phosphatidylethanolamines; D010743:Phospholipids; C043248:mercury-dextran; D019856:Ethanolamine; D014302:Trinitrobenzenesulfonic Acid; D002784:Cholesterol; D009251:NADPH-Ferrihemoprotein Reductase; D003576:Electron Transport Complex IV; D011402:Pronase; D000254:Sodium-Potassium-Exchanging ATPase; D008628:Mercury