O-fucosylation of thrombospondin type 1 repeats.
Thrombospondin type 1 repeats (TSRs) are small cysteine-rich motifs with three conserved disulfide bonds originally described as modules in the thrombospondins. Since then, TSRs have been found as tandem repeats in a wide variety of secreted and cell-surface proteins of diverse function. TSRs in many contexts are known to bind a variety of receptors and have antiangiogenic capabilities. They can be modified with O-linked fucose on serine/threonine found in the consensus, CX(2-3)(S/T)CX(2)G. Here we review what is known about O-fucosylation of TSRs and describe in detail mass spectral methods to map sites of O-fucosylation on proteins containing TSRs. These methods include techniques to identify glycosylated peptides and the relative amounts of elongated products by electrospray ionization mass spectrometry of glycopeptides.
Leonhard-Melief
Christina
C
Department of Biochemistry and Cell Biology, Institute for Cell and Developmental Biology, Stony Brook University, Stony Brook, New York, USA.
Haltiwanger
Robert S
RS
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D002853:Chromatography, Liquid / Q000379:methods; D005643:Fucose / Q000378:metabolism*; D006031:Glycosylation; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D010449:Peptide Mapping / Q000379:methods; D011499:Protein Processing, Post-Translational; D020449:Repetitive Sequences, Amino Acid; D019699:Thrombospondins / Q000737:chemistry* / Q000378:metabolism*
D019699:Thrombospondins; D005643:Fucose