- Volume 1 Issue 1
A Simple Purification of Apoliproteins A-I and B and Their Application to Cholestery Ester Transfer Assay
- Cho, Kyung-Hyun (Dept. of Genetic Engineering, Kyungpook national University, Taegu 702-701) ;
- Park, Myung-Sook (Dept. of Food Science and Nutrition) ;
- Bok, Song-Hae (Bioproducts Research Group, Genetic Engineering Research Institute, KIST, Taejon 305-606) ;
- Park, Young-Bok (Dept. of Genetic Engineering, Kyungpook national University, Taegu 702-701)
- Published : 1996.06.01
This study describes a stable and simple method for the measurement of cholesteryl ester transfer protein(CETP) activities using reconstituted HDL and LDL as substrates. Apolipoproteins (apo) A-I and -B were purified from hog plasma by a new strategy without ultracentrifugation and delipidation. a simple two-step column chromatography was administered. In the first step of phenyl-sepharose CL-4B column chro-matography, hydrophobic plasma proteins were isolated. The most hydrophobic proteins bound to the column appeared to be A-I and apo-B. Contaminat proteins were efficiently eliminated from the sample by washing the column with 0.3M NaCI containing buffer after loading the plasma on the column. Two pure proteins showing each single band on SDS-PSGE of apo A-I and apo-B were individually obtained by a subsequent gel filtration column chromatography(Sephadex G-200). This two-step purification was simple and inexpensive compared to the ultracentrifugation and/or delipidation method that are most commonly used. Reconstituted hight-density lipoproteins(HDL) and low-density lipoproteins(LDL) were prepared using the purified apo A-I and-B, respectively. When these artificially prepared HDL and LDL were used in the assays for CETP as the cholesteryl ester(CE) donor and acceptor respectively, the specific transfer of CE increased up to two fold compared to that used the native HSL and LDL.
- J. Lipid Res. v.34 Plasma cholesteryl ester transfer protein Tall,A.R.
- J. Clin. Invest. v.86 Plasma high density lipoproteins Metabolism and relationship to atherogenesis Tall,A.R.
- Atherosclerosis v.58 Deficiency of serum cholesteryl-ester transfer activity in patients with familial hyperalphalipoproteinemia Koizumi,J.;Mabuchi,H.;Yoshimura,A.;Michishita,I.;Takeda,M.;Itoh,H.;Sakai,T.;Nuda,K.;Takeda,R.
- Artery. v.14 High density lipoprotein with poor reactivity to cholesteryl ester transfer reaction observed in a homozygote of familial hyperalphalipoproteinemia Yokoyama,S.;Kurasawa,T.;Nishikawa,O.;Yamamoto,A.
- Diabetes/Metabolism Reviews v.7 The role of the cholesteryl ester transfer protein in lipoprotein metabolism Swenson,T.L.
- Clin. Chim. Acta. v.178 Apolipoprotein measurements in clinical biochemistry and their utility visa-vis conventional assays Bachorik,P.S.;Kwiterovih,P.O.Jr.
- J. Am. Med. Assoc. v.251 Lipid Research Clinics Program: The lipid research clinics coronary primary prevention trial results. Ⅰ. Reduction in incidence of coronary artery diseases
- J. Am. Med. Assoc. v.251 Lipid Research Clinics Program: The lipid research clinics coronary primary prevention trial results. Ⅱ. The relationship of reduction in incidence of coronary artery diseases to cholesterol lowering
- Clin. Chem. v.33 Cholesteryl ester transfer activity in plasma measured by using solid-phase-bound high-density lipoprotein Sparks,D.L.;Frohlich,J.;Cullis,P.;Pritchard,P.H.
- J. Biol. Chem. v.265 A plasma inhibitor of triglyceride and cholesteryl ester transfer activities Morton,R.E.;Zilversmit,D.B.
- Clin. Chem. v.32 Improved estimation of cholesteryl ester transfer/exchange activity in serum or plasma Groener,J.E.M.;Pelton,R.W.;Kostner,G.M.
- J. Clin. Invest. v.34 The distribution and chemical composition of ultracentrifugally separated lipoproteins in human serum Havel,R.J.;Eder,H.A.;Bragdon,J.H.
- J. Lipid Res. v.17 A solvent system for delipidation of plasma or serum without protein precipitation Cham,B.E.;Knowles,B.R.
- J. Biol. Chem. v.248 A comparison of the major apolipoprotein from pig and human high density lipoproteins Richard,L.J.;Baker,N.H.;Tauton,O.D.;Smith,L.C.;Garner,C.W.;Gotto,A.M.Jr.
- Nature (London) v.227 Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Laemmli,U.K.
- Acta. Pathol. Microbiol. Scand. v.26 Antigen-antibody reactions in gels Ouchterlony,O.
- Biochim. Biophys. Acta v.231 Single bilayer liposomes prepared without sonication Batzri,S.;Korn,E.D.
- J. Lipid Res. v.23 Characterization of proteoliposomes containing apolipoprotein A-1 : A new substrate for the measurement of lecithin : cholesterol acyltranferase activity Chen,C.H.;Albers,J.J.
- J. Lipid Res. v.31 Impaired plasma cholesteryl ester transfer with accumulation of large high density lipoproteins in some families of baboons(Papio sp.) Kushwaha,R.S.;Rainwater,D.L.;Wiliams,M.C.;Getz,G.S.;McGill,H.C.;Jr.
- J. Lipid Res. v.4 Characterization of cholesteryl ester transfer protein inhibitor from plasma of baboons(Papio sp.) Kushwaha,R.S.;Hasan,S.Q.;McGill,H.C.Jr.;Getz,G.S.;Dunham,R.G.;Kanda,P.
- Anal. Biochem. v.72 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Bradford. M.M.
- Kor. Biochem. J. v.25 Reaction of HDL-bound cholesteryl ester traosfer protein and a new method for its activity measurement Park,Y.B.;Jeoung,N.H.;Kim,H.S.;Choi,M.S.
- Med. Clin. N. Am. v.66 Cholesterol transport between cells and body fluids Fielding,C.J.;Fielding,P.E.