Immunological characterization of neutral cholesteryl ester hydrolase from rat liver cytosol.
Rabbit polyclonal antibodies were raised against rat liver bile salt-independent neutral cholesteryl ester hydrolase (CEH) and used for subcellular localization and immunological comparison with isoforms from other tissues. Antibodies exhibited a high degree of specificity for the liver CEH through all stages of purification and neutralized 70-80% of the activity of liver cytosolic CEH. They exhibited various levels of cross-reactivity with cytosolic proteins from other tissues, but reacted weakly with pancreatic and intestinal proteins and did not inhibit pancreatic CEH. Cytosol contained 78% of total cellular CEH activity and 75% of CEH immunoreactive protein. Washed microsomes contained 3% of CEH activity and 5% of CEH protein.
Ghosh
S
S
Department of Biochemistry and Molecular Biophysics, Medical College of Virginia, Virginia Commonwealth University, Richmond 23298-0614.
Grogan
W M
WM
D000818:Animals; D000918:Antibody Specificity / Q000276:immunology; D015153:Blotting, Western; D003429:Cross Reactions / Q000276:immunology; D003600:Cytosol / Q000201:enzymology*; D004797:Enzyme-Linked Immunosorbent Assay; D008099:Liver / Q000201:enzymology*; D051381:Rats; D002787:Sterol Esterase / Q000032:analysis*; D013347:Subcellular Fractions / Q000201:enzymology
D002787:Sterol Esterase