Highly sensitive sequencing of the sulfated domains of heparan sulfate.
The heparan sulfates (HS) are hypervariable linear polysaccharides that act as membrane co-receptors for growth factors, chemokines, and extracellular matrix proteins. In most instances, the molecular basis of protein recognition by HS is poorly understood. We have sequenced 75% of the sulfated domains (S-domains) of fibroblast HS, including all of the major ones. This analysis revealed tight coupling of N- and 2-O-sulfation and a low frequency but precise positioning of 6-O-sulfates, which are required functional groups for HS-mediated activation of the fibroblast growth factors. S-domain sequencing was conducted using a novel and highly sensitive method based on a new way of reading the sequence from high performance liquid chromatography separation profiles of metabolically labeled HS-saccharides following specific chemical and enzymatic scission. The implications of the patterns seen in the sulfated domains for better understanding of the synthesis and function of HS are discussed.
Merry
C L
CL
Cancer Research Campaign and University of Manchester Department of Medical Oncology, Christie Hospital NHS Trust, Manchester, M20 4BX, United Kingdom.
Lyon
M
M
Deakin
J A
JA
Hopwood
J J
JJ
Gallagher
J T
JT
D016475:3T3 Cells; D000818:Animals; D002240:Carbohydrate Sequence; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D006497:Heparan Sulfate / Q000737:chemistry*; D051379:Mice; D008969:Molecular Sequence Data; D013431:Sulfates / Q000737:chemistry*
D006497:Heparan Sulfate; D013431:Sulfates