Identification and characterization of recombinant murine interleukin-6 with a C-terminal pentapeptide extension using capillary reversed phase HPLC-MS and Edman degradation
- Author(s)
- Hammacher, A; Reid, GE; Moritz, RL; Simpson, RJ;
- Details
- Publication Year 1997-11,Volume 11,Issue #6,Page 337-342
- Journal Title
- BIOMEDICAL CHROMATOGRAPHY
- Publication Type
- Journal Article
- Abstract
- We have identified a preparation of recombinant murine interleukin-6 (mIL-6) that, in addition to the anticipated product, also contained approximately equal amounts of mIL-6 with a C-terminal pentapeptide extension, The extension mutant was generated by readthrough of the stopcodon, and termination at a second in-frame stopcodon 12 base pairs 3' in the expression vector. Aliquots of the preparation were subjected to proteolytic digestion with Asp-N and Lys-C-endopeptidase, The resultant peptides were separated by reversed-phase capillary HPLC, and analysed using a combination of mass spectrometry and N-terminal sequence analysis, These data revealed a C-terminal pentapeptide (Gln-Gly-Ser-Val-Asp) extension, with the authentic stopcodon being translated as glutamine, The extension mutant was isolated by reversed-phase HPLC and shown to have similar mitogenic activity to mIL-6 on murine hybridoma 7TD1 cells. (C) 1997 John Wiley & Sons, Ltd.
- Publisher
- JOHN WILEY & SONS LTD
- Keywords
- ESCHERICHIA-COLI; SYNTHETIC GENE; PURIFICATION; GROWTH; IL-6; METHIONINE; EXPRESSION; PROTEINS; RECEPTOR; SEQUENCE
- Publisher's Version
- https://doi.org/10.1002/(SICI)1099-0801(199711)11:6<337::AID-BMC687>3.3.CO;2-5
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- Refer to copyright notice on published article.
Creation Date: 1997-11-01 12:00:00