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Remodeling of the lectin–EGF-like domain interface in P- and L-selectin increases adhesiveness and shear resistance under hydrodynamic force

Nature Immunology volume 7, pages 883–889 (2006)Cite this article

Abstract

Crystal structures of the lectin and epidermal growth factor (EGF)–like domains of P-selectin show 'bent' and 'extended' conformations. An extended conformation would be 'favored' by forces exerted on a selectin bound at one end to a ligand and at the other end to a cell experiencing hydrodynamic drag forces. To determine whether the extended conformation has higher affinity for ligand, we introduced an N-glycosylation site to 'wedge open' the interface between the lectin and EGF-like domains of P-selectin. This alteration increased the affinity of P-selectin for its ligand P-selectin glycoprotein 1 (PSGL-1) and thereby the strength of P-selectin-mediated rolling adhesion. Similarly, an asparagine-to-glycine substitution in the lectin-EGF-like domain interface of L-selectin enhanced rolling adhesion under shear flow. Our results demonstrate that force, by 'favoring' an extended selectin conformation, can strengthen selectin-ligand bonds.

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Figure 1: Selectin variants.
Figure 2: Wedge mutant P-selectin has increased adhesiveness to HL-60 cells.
Figure 3: Rolling and transient tethering of wedge mutant P-selectin.
Figure 4: Increased affinity of wedge mutant P-selectin.
Figure 5: Increased binding of wedge mutant soluble P-selectin to PSGL-1 transfectants.
Figure 6: Enhancement of rolling adhesiveness by the N138G substitution in L-selectin.
Figure 7: The N138G substitution in L-selectin increases accumulation in shear flow and decreases transient tether koff.

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Acknowledgements

We thank R. Camphausen for suggestions and discussion. Supported by Kirschstein National Research Service Award (F32 AI052851 to U.T.P., F32 GM073529 to T.T.W. and HL-48675 to T.A.S).

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  1. Uyen T Phan

    Present address: Organon, Cambridge, Massachusetts, 02142, USA

Authors and Affiliations

  1. Department of Pathology, CBR Institute for Biomedical Research, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Uyen T Phan, Travis T Waldron & Timothy A Springer

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Correspondence to Timothy A Springer.

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Supplementary information

Supplementary Table 1

Primers were used in PCR-based mutagenesis to generate the various P-selectin and L-selectin variants. (PDF 49 kb)

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Phan, U., Waldron, T. & Springer, T. Remodeling of the lectin–EGF-like domain interface in P- and L-selectin increases adhesiveness and shear resistance under hydrodynamic force. Nat Immunol 7, 883–889 (2006). https://doi.org/10.1038/ni1366

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