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Release of low density lipo protein receptors from human fibroblasts of hela cells by tryptic digestion



Release of low density lipo protein receptors from human fibroblasts of hela cells by tryptic digestion



Biochemical and Biophysical Research Communications 84(2): 366-373



Trypsin treatment of cultured normal human skin fibroblasts or HeLa (human cervical cancer) 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 6-[3H]-glucosamine .cntdot. HCl and [3H]-leucine. It is inactivated by heating at 80.degree. 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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Accession: 006306071

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PMID: 214072



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