Purification and partial amino acid sequence analysis of two distinct tumor necrosis factor receptors from HL60 cells.

摘要: Two distinct tumor necrosis factor (TNF) receptors of 55- and 75-kDa apparent molecular masses previously identified on the cell surface by monoclonal antibodies have been solubilized with Triton X-100 from HL60 cells. A filter-based dot blot assay was developed to monitor specific 125I-TNF alpha binding during fractionation extract. By a combination immuno- ligand affinity chromatography reverse phase high performance liquid both receptor proteins were purified homogeneity. Analysis sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed two bands at 55 51 kDa for 55-kDa TNF major minor 65-kDa band receptor. All these specifically bound beta in experiments. The exclusive specificity utr series 75.65-kDa htr 55.51-kDa demonstrated antigens Western blots. Both types found contain N-linked carbohydrates. N-terminal amino acid sequence analysis 51-kDa revealed identical sequences suggesting possible truncation C-terminal end. different determined band. One corresponded published ubiquitin; other therefore assumed be unique Additional internal this after proteolytic cleavage.