C-Mannosylation andO-Fucosylation of Thrombospondin Type 1 Repeats

Anne Gonzalez de Peredo (1) , Dominique Klein , Boris Macek , Daniel Hess , Jasna Peter-Katalinic , Jan Hofsteenge (1)

1 FMI - Friedrich Miescher Institute for Biomedical Research

The final chemical structure of a newly synthesized protein is often only attained after further covalent modification. Ideally, a comprehensive proteome analysis includes this aspect, a task that is complicated by our incomplete knowledge of the range of possible modifications and often by the lack of suitable analysis methods. Here we present two recently discovered, unusual forms of protein glycosylation, i.e. C-mannosylation and O-fucosylation. Their analysis by a combined mass spectrometric approach is illustrated with peptides from the thrombospondin type 1 repeats (TSRs) of the recombinant axonal guidance protein F-spondin. Nano-electrospray ionization tandem-mass spectrometry of isolated peptides showed that eight of ten Trp residues in the TSRs of F-spondin are C-mannosylated. O-Fucosylation sites were determined by a recently established nano-electrospray ionization quadrupole time-of-flight tandem-mass spectrometry approach. Four of five TSRs carry the disaccharide Hex-dHex-O-Ser/Thr in close proximity to the C-mannosylation sites. In analogy to thrombospondin-1, we assume this to be Glc-Fuc-O-Ser/Thr. Our current knowledge of these glycosylations will be discussed.

Mots clés: MS; mass spectrometry; CID; collision-induced dissociation; MSMS; tandem-mass spectrometry; Q-TOF; quadrupole time-of-flight; rF-spondin; recombinant F-spondin; TSP-1; thrombospondin-1; TSR; thrombospondin type 1 repeat; ESI; electrospray ionization; EGF; epidermal growth factor; LC; liquid chromatography; PMP-C; pars intercerebralis major peptide C

Domaines: Sciences du Vivant [q-bio]

Anne Gonzalez de Peredo, Dominique Klein, Boris Macek, Daniel Hess, Jasna Peter-Katalinic, et al.. C-Mannosylation andO-Fucosylation of Thrombospondin Type 1 Repeats. Molecular and Cellular Proteomics, 2001, 1 (1), pp.11 - 18. ⟨10.1074/mcp.m100011-mcp200⟩. ⟨hal-03080321⟩