ABSTRACT
Selectins are a family of adhesion molecules with a well-defined domain structure comprised of a lectin or carbohydrate-recognition domain (CRD), an epidermal growth factor (EGF)-like motif, and a variable number of consensus repeats (CRs). While it is clear from various lines of evidence that the CRD plays a pivotal role in selectin-ligand interactions, little is known about the role of the non-lectin selectin domains. We expressed a series of soluble chimeric proteins with various domains switched between E- and L-selectin and measured binding of the resulting chimeras to sialyl Lewis(a) and sulfatide, two carbohydrate structures which are specific for the E- and L-CRDs, respectively. Both CRDs bind to their respective ligands with the same affinity regardless of the origin of the other domains they are attached to. The domain-switched chimeras were assayed for their ability to support static binding and rolling of various cell lines which bind specifically to E-selectin. In these assays, the E-CRD was indispensable for both static binding and rolling under physiological flow conditions. The E-CRD alone, when substituted into L-selectin, supported rolling without the requirement for additional ligand-recognition elements. We conclude that the EGF domain or the CRs of E- and L-selectin have no influence on the CRD's specificity to carbohydrates. Furthermore, at least in the case of E-selectin, they do not contribute to the specificity of binding to cell surface ligands.
