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. Copyright (c) 2010 Elsevier Inc. All rights reserved.