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Papers In Press, published online ahead of print November 5, 2003
J. Biol. Chem, 10.1074/jbc.M311398200
Submitted on October 17, 2003
Revised on November 4, 2003
Accepted on November 4, 2003

Determining heparan sulfate structure in the vicinity of specific sulfotransferase recognition sites by mass spectrometry

Zhengliang L. Wu, Miroslaw Lech, David L. Beeler, and Robert D. Rosenberg

Biology, Massachusetts Institute of Technology, Cambridge, MA 02142

Corresponding Author: rdrrosen{at}MIT.EDU

Sulfated motifs on heparan sulfate (HS) are involved in various extracellular processes from cell signaling to enzymatic regulation, but the structures of these motifs are obscure. We have developed a strategy to determine the structure of sulfotransferase recognition sites which constitute these motifs. Stable isotope is first introduced into specific sites on HS with HS sulfotransferases and the modified HS is then digested into oligosaccharides of differing sizes. The overlapping oligosaccharides containing the introduced stable isotope are identified by changes in the m/z profiles by mass spectrometry, and their relationships are elucidated. In this way, the HS structures in the vicinity of the sulfotransferase recognition site are quickly determined and groups on precursor structures of HS that direct the action of HS sulfotransferases are pinpointed.


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