Release of low density lipoprotein receptors from human fibroblasts or HeLa cells by tryptic digestion. |
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Authors: | N Di Ferrante P V Donnelly D Di Ferrante S Toma A M Gotto |
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Institution: | 1. Laboratories of Connective Tissue Research of the Department of Biochemistry, Baylor College of Medicine, Houston, Texas 77030 USA;2. Division of Atherosclerosis and Lipoproteins of the Department of Medicine, Baylor College of Medicine, Houston, Texas 77030 USA;3. The Methodist Hospital Houston, Texas 77030 USA |
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Abstract: | Trypsin treatment of cultured normal human skin fibroblasts or Hela cells releases material which is retained on a low density lipoprotein (LDL)-Sepharose affinity column, may be eluted from it with 2.5 M KI and, after dialysis, agglutinates LDL or apo-B-coated formocells. Such agglutination is prevented by preincubation of the receptor with LDL in solution or with arginine-rich protamine. Trypsin treatment of “receptor defective” or “receptor negative” mutant fibroblasts releases material which is retained on LDL-Sepharose column but fails to agglutinate LDL-coated formocells. The receptor may be labeled with and ]-leucine, it is inactivated by heating at 80°C for 10 min and may be obtained from normal fibroblasts or Hela cells, whether they were cultured in presence or in absence of lipoprotein-containing fetal calf serum. |
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Keywords: | LDL low density lipoproteins apo-B apolipoprotein B LDS lipoprotein deficient serum FCS fetal calf serum |
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