Recombinant-derived interleukin-1{alpha} stabilized against specific deamidation |
| |
Authors: | Wingfield Paul T; Mattaliano Robert J; MacDonald HRobson; Craig Stewart; Clore GMarius; Gronenborn Angela M; Schmeissner Ursula |
| |
Affiliation: | Biogen SA PO Box 1211 Geneva, Switzerland
1Biogen Research Corporation 14 Cambridge Center, Cambridge, MA 02142, USA
2Ludwig Institute for Cancer Research, Lausanne Branch 1066 Epalinges, Switzerland
3Department of Biochemistry, University of Newcastle Newcastle upon Tyne NE1 7RU, UK
4Max-Planck-institut fur Biochemuie 8033 Msrtinsried, FRG |
| |
Abstract: | Recombinant-derived human interleukln-1 (IL-1 ), purified fromEscherichia coli, was resolved by isoelectric focusing on polyacrylamidegels into two species of isoelectric points (pI) 5.45 and 5.20,which constituted 75% and 25% of the total IL-1 protein respectively.The pI 5.45 and pI 5.20 species were separated by chromatofocusingand subjected to N-terminal sequence analysis. The pI 5.45 speciescontained the expected Asn residue at position 36 of the matureprotein sequence whereas the pI 5.20 species contained an Aspresidue at the same position. A mutant protein in which Asn-36was substituted for a Ser residue was isolated from E.coli andshown to be homogeneous on isoelectric focusing analysis witha pI = 5.45. 1H-n.m.r. and circular dichroism analyses of wild-typeand the mutant IL-1 indicated a similar conformation which wasalso indicated by the identical receptor binding affinitiesof IL-1 with Asn, Asp or Ser in position 36. The mutant proteinwas stabilized against specific base catalysed and temperature-induceddeamidation, and may be more suitable than the wild-type positionfor physical and structural studies. |
| |
Keywords: | interleukin-1 /" target="_blank">gif" ALT="{alpha}" BORDER="0">/ sequence determmation/ deamidanon/ site-specific mutagenesis/ protein conformation |
本文献已被 Oxford 等数据库收录! |
|