O-fucosylation of thrombospondin type 1 repeats

Methods Enzymol. 2010;480:401-16. doi: 10.1016/S0076-6879(10)80018-7.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Review

MeSH terms

  • Amino Acid Motifs
  • Amino Acid Sequence
  • Animals
  • Catalytic Domain
  • Chromatography, Liquid / methods
  • Fucose / metabolism*
  • Glycosylation
  • Humans
  • Mass Spectrometry / methods
  • Peptide Mapping / methods
  • Protein Processing, Post-Translational
  • Repetitive Sequences, Amino Acid*
  • Thrombospondins / chemistry*
  • Thrombospondins / metabolism*

Substances

  • Thrombospondins
  • Fucose