Purification and identification of two distinct isoforms of rabbit pancreatic cholesterol esterase.
Cholesterol esterase (CEases; E.C. 3.1.13) has been purified to homogeneity from rabbit pancreas. The method of purification consists of homogenization of total pancreas, high speed centrifugation, anion exchange column chromatography on S-Sepharose, size exclusion on Sephacryl followed by affinity chromatography on heparin agarose. During the purification procedure, two distinct isoforms of CEases have been identified. Both forms are similar in their molecular weights, bile salt requirement and pH optima but differ in their sensitivity to heparin. Isoform-I is resistant and isoform-II is sensitive to heparin. In the normal pancreas of the adult rabbit, the amount of each of the enzymes appears to be in equimolar concentrations. Physiological significance of the existence of heparin sensitive and resistant forms by the same tissue is unclear. In view of the significant role played by heparin in the modulation of CEase activity and several other physiological functions, these two isoforms may have different mechanisms of action on the hydrolysis of carboxyl esters of cholesterol and vitamins.
Jamry
W
W
Division of Geriatric Research Education and Clinical Center, VA Medical Center, St. Louis, MO 63125, USA.
Sasser
T
T
Kumar
B V
BV
D000818:Animals; D001647:Bile Acids and Salts / Q000378:metabolism; D006493:Heparin / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000187:drug effects / Q000302:isolation & purification* / Q000378:metabolism; D010179:Pancreas / Q000201:enzymology*; D011817:Rabbits; D002787:Sterol Esterase / Q000187:drug effects / Q000302:isolation & purification* / Q000378:metabolism*
D001647:Bile Acids and Salts; D007527:Isoenzymes; D006493:Heparin; D002787:Sterol Esterase