We report a simple method, designated "spot transfer", where known proteins are excised and eluted from a two-dimensional (2-D) gel run on one gel system to transfer identification of the proteins to a different 2-D gel system by comigration with a comparable sample. In one experiment, 8 of 16 proteins eluted from an isoelectric focusing (IEF) 2-D gel, of the format described for the Celis human keratinocyte database (Celis, J. E. et al., Electrophoresis 1993, 14, 1091-1198), were found to comigrate with proteins in a human T lymphoma (JURKAT) whole cell lysate run using the Millipore Investigator 2-D system. The method should have general utility in allowing the exchange of protein identifications between investigators and could be used to standardize gel loci used in 2-D gel protein databases.
A biologically active preparation of murine recombinant interleukin‐1β (mIL‐1β) fromEscherichia coli cell lysates contained two forms of mIL‐1β with pI 8.7 and pI 8.1, respectively. Treatment with 0.1 M Tris, pH 8.5, at 37°C for 35 h converted the pI 8.7 form to the pI 8.1 form by the selective deamidation of an asparagine residue (Asn149) in the mIL‐1β molecule. Deamidated mIL‐1β had 3‐ to 5‐fold lower co‐mitogenic activity and receptor affinity than the unmodified form.
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