Cholesterol-transfer protein located in the intestinal brush-border membrane. Partial purification and characterization.
Cholesterol absorption by small intestinal brush border membrane vesicles from taurocholate mixed micelles is a second-order reaction. From a comparison of reaction rates and order before and after proteinase K treatment of brush-border membrane vesicles, it is concluded that cholesterol absorption is protein-mediated. It is shown that the desorption of cholesterol from taurocholate mixed micelles is by a factor of about 10(4) faster than that from egg phosphatidylcholine bilayers. When brush border membrane vesicles are stored at room temperature, intrinsic proteinases are activated and proteins are liberated from the brush border membrane. These proteins collected in the supernatant catalyze cholesterol and phosphatidylcholine exchange between two populations of small unilamellar phospholipid vesicles. One of the active proteins present in the supernatant is purified by a two-step procedure involving gel filtration on Sephadex G-75 SF and affinity chromatography on a Nucleosil-phosphatidylcholine column. The protein thus obtained is pure by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. It has an apparent molecular weight of slightly less than 14,000 as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis and a value of 11,500 determined by gel filtration on Sephadex G-75 SF.
Thurnhofer
H
H
Laboratorium für Biochemie, Eidgenössische Technische Hochschule Zürich, Switzerland.
Schnabel
J
J
Betz
M
M
Lipka
G
G
Pidgeon
C
C
Hauser
H
H
D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry / Q000302:isolation & purification*; D002784:Cholesterol / Q000378:metabolism; D053480:Cholesterol Ester Transfer Proteins; D002845:Chromatography; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins; D006868:Hydrolysis; D007408:Intestinal Absorption; D007422:Intestines / Q000737:chemistry* / Q000648:ultrastructure; D007700:Kinetics; D008244:Lysophosphatidylcholines; D008823:Micelles; D008871:Microvilli / Q000737:chemistry*; D011817:Rabbits; D013656:Taurocholic Acid
D002352:Carrier Proteins; D053480:Cholesterol Ester Transfer Proteins; D006023:Glycoproteins; D008244:Lysophosphatidylcholines; D008823:Micelles; D013656:Taurocholic Acid; D002784:Cholesterol