Effects of sulfate position on heparin octasaccharide binding to CCL2 examined by tandem mass spectrometry.
Chemokine-glycosaminoglycan (GAG) interactions have been shown to be essential for in vivo chemokine signaling, which functions in such diverse processes as inflammation, development, and cancer metastasis. Despite the importance of these interactions, the saccharide sequence dependency of chemokine-GAG interactions is poorly understood. In a recent study, FT-ICR mass spectrometry was used to show that the chemokine CCL2 (monocyte chemoattractant protein 1) binds only to the 11- and 12-sulfated components of a heparin octasaccharide library. Although the exact structure of the fully sulfated, 12-sulfated octasaccharide is known, the 11-sulfated species could have a number of sulfated disaccharide sequences. In the current study, the composition of the 11-sulfated heparin octasaccharides, as well as the composition of CCL2 affinity purified 11-sulfated heparin octasaccharides, were examined by tandem MS. Of the three possible singly desulfated disaccharides, one species, III-S, is enriched by CCL2 affinity purification, indicating that the 11-sulfated heparin octasaccharides containing this disaccharide are preferentially bound to CCL2. These data suggest that 2-O and N sulfation of heparin may be of greater importance to CCL2-heparin binding than 6-O sulfation.
Sweeney
Matt D
MD
Genome Center, Department of Chemistry, Room 1300, University of California, 1 Shields Avenue, 95616, Davis, CA, USA.
Yu
Yonghao
Y
Genome Center, Department of Chemistry, Room 1300, University of California, 1 Shields Avenue, 95616, Davis, CA, USA.
Leary
Julie A
JA
Genome Center, Department of Chemistry, Room 1300, University of California, 1 Shields Avenue, 95616, Davis, CA, USA. jaleary@ucdavis.edu.
D001665:Binding Sites; D018932:Chemokine CCL2 / Q000737:chemistry*; D006493:Heparin / Q000737:chemistry*; D009844:Oligosaccharides / Q000737:chemistry*; D011485:Protein Binding; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*; D013329:Structure-Activity Relationship; D013431:Sulfates / Q000737:chemistry*
D018932:Chemokine CCL2; D009844:Oligosaccharides; D013431:Sulfates; D006493:Heparin