Identification of an E-selectin region critical for carbohydrate recognition and cell adhesion.
E-selectin elicits cell adhesion by binding to the cell surface carbohydrate, sialyl Lewis X (sLe(x)). We evaluated the effects of mutations in the E-selectin lectin domain on the binding of a panel of anti-E-selectin mAbs and on the recognition of immobilized sLe(x) glycolipid. Functional residues were then superimposed onto a three-dimensional model of the E-selectin lectin domain. This analysis demonstrated that the epitopes recognized by blocking mAbs map to a patch near the antiparallel beta sheet derived from the NH2 and COOH termini of the lectin domain and two adjacent loops. Mutations that affect sLe(x) binding map to this same region. These results thus define a small region of the E-selectin lectin domain that is critical for carbohydrate recognition.
Erbe
D V
DV
Department of Immunology, Genentech, Inc., South San Francisco, California 94080.
Wolitzky
B A
BA
Presta
L G
LG
Norton
C R
CR
Ramos
R J
RJ
Burns
D K
DK
Rumberger
J M
JM
Rao
B N
BN
Foxall
C
C
Brandley
B K
BK
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001665:Binding Sites / Q000502:physiology; D002448:Cell Adhesion / Q000502:physiology*; D015815:Cell Adhesion Molecules / Q000737:chemistry / Q000378:metabolism*; D004252:DNA Mutational Analysis; D019040:E-Selectin; D005455:Fluorescent Antibody Technique; D006801:Humans; D016256:Lewis X Antigen / Q000737:chemistry / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011817:Rabbits; D011971:Receptors, Immunologic / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D000911:Antibodies, Monoclonal; D015815:Cell Adhesion Molecules; D019040:E-Selectin; D016256:Lewis X Antigen; D008562:Membrane Glycoproteins; D011971:Receptors, Immunologic; D011993:Recombinant Fusion Proteins