title: Recombinant human interleukin-12 is the second example of a C-mannosylated protein.
abstract: The beta-chain of human interleukin 12 (IL-12) contains at position 319-322, the sequence Trp-x-x-Trp. In human RNase 2 this is the recognition motif for a new, recently discovered posttranslational modification, i.e., the C-glycosidic attachment of a mannosyl residue to the side chain of tryptophan. Analysis of C-terminal peptides of recombinant IL-12 (rHuIL-12) by mass spectrometry and NMR spectroscopy revealed that Trp-319beta is (partially) C-mannosylated. This finding was extended by in vitro mannosylation experiments, using a synthetic peptide derived from the same region of the protein as an acceptor. Furthermore, human B-lymphoblastoid cells, which secrete IL-12, were found to contain an enzyme that carries out the C-mannosylation reaction. This shows that nonrecombinant IL-12 is potentially C-mannosylated as well. This is only the second report on a C-mannosylated protein. However, the occurrence of the C-mannosyltransferase activity in a variety of cells and tissues, and the presence of the recognition motif in many proteins indicate that more C-mannosylated proteins may be found.
authors: Friedrich Miescher-Institut, P.O. Box 2543, CH-4002 Basel, Switzerland.
authors: M A
authors: MA
authors: Doucey
authors: D
authors: D
authors: Hess
authors: M J
authors: MJ
authors: Blommers
authors: J
authors: J
authors: Hofsteenge
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001402:B-Lymphocytes / Q000378:metabolism; D016466:CHO Cells; D006224:Cricetinae; D006031:Glycosylation; D006801:Humans; D066298:In Vitro Techniques; D018664:Interleukin-12 / Q000737:chemistry* / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008358:Mannose / Q000737:chemistry* / Q000378:metabolism; D008364:Mannosyltransferases / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D010446:Peptide Fragments; D011994:Recombinant Proteins; D018664:Interleukin-12; D008364:Mannosyltransferases; D008358:Mannose