CHARACTERIZATION OF THE 14 KDA FRAGMENT OF HUMAN TUMOR-NECROSIS-FACTOR-ALPHA
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- Published online on: December 1, 1995 https://doi.org/10.3892/ijo.7.6.1437
- Pages: 1437-1443
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Abstract
We report the characterization of a 14 kDa degradation fragment from recombinant human tumour necrosis factor-alpha (TNF alpha) by N-terminal sequencing and mass spectrometry. A single site between the dibasic residues Arg(31)-Arg(32) of the mature recombinant 17 kDa protein has been identified as the target site that generates the 14 kDa fragment. The observation that a maximum of 33% degradation occurs suggests that only one monomer per TNF trimer is cleaved. E. coli proteases specific for dibasic residues are thought to be responsible for this cleavage. A strategy has been developed which completely inhibits proteolysis. This strategy has been used to reduce the 14 kDa degradation fragment obtained from approximately 33% of the total purified protein to zero.
