FIGURE 1.

Sequence analysis of HS^NS4F5. A, affinity chromatography on NS4F5-loaded columns using size-defined, ³H end-labeled heparin oligosaccharides (4–12-mer) and a 12-mer oligosaccharide preparation affinity selected on antithrombin III (12-mer HA). B, fractionation of NS4F5 affinity-purified octasaccharides (as in A) before (upper panel) or after (lower panel) partial deaminative cleavage by anion exchange HPLC on a Propac column, using a linear salt gradient for elution (diagonal line). C, reactivity of immobilized NS4F5 with heparin-derived, ³H end-labeled octasaccharide libraries with defined numbers of 2-O-sulfate groups and variable numbers of 6-O-sulfate groups using a stepwise increasing salt gradient as indicated (dotted line). The left column shows affinity separation of oligosaccharides with two 2-O-sulfate groups and zero, one, two, or three added 6-O-sulfate groups, respectively. The right panel displays corresponding patterns for octasaccharides containing three 2-O-sulfate groups (n = 4).