Journal of Biological Chemistry · 1980 · 238 citations · 34 references
Life SciencesBiochemistryMolecular BiologyEnzymologyLipid BiochemistrySteroid BiochemistryCholesterol BiosynthesisCholesterol MetabolismLipid MetabolismSteroid BiosynthesisSteroid MetabolismSterol MetabolismGel ElectrophoresisHepatic Sterol MetabolismIsoelectric FocusingLanosterol MetabolismMolecular Weight DeterminationPolyacrylamide Gel ElectrophoresisPurificationSodium Dodecyl Sulfate-polyacrylamide Gel Electrophoresis
The present article describes the purification and properties of sterol carrier proteinz (SCPz), which has been purified to apparent homogeneity from rat liver 303,000 X g soluble supernatant ( s 3 0 3 ) .SCPZ activates the microsomal conversion of 4,4-dimethyl-A8-cholestenol to CZ, sterol and the microsomal conversion of 7dehydrocholesterol to cholesterol.The purification factors achieved during SCPZ purification were essentially the same (1400-to 1500-fold) for both substrates.In contrast to SCPl, SCPZ did not activate the microsomal conversion of squalene to lanosterol.Evidence for the homogeneity of the purified SCPz includes the following: (i) a single band in basic, acidic, and sodium dodecyl sulfate polyacrylamide gel electrophoresis, (ii) a single band in analytical isoelectric focusing, and (iii) exact correspondence between SCPz activity and stained protein absorbance in polyacrylamide gel electrophoresis.SCPz is a basic protein (PI E 8.6) with a molecular weight of 13,500, as measured by sodium dodecyl sulfate polyacrylamide gel electrophoresis.The most abundant amino acid is lysine (14.0 mole 8).In addition, SCP2 has no arginine or tyrosine.
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