Effect of lengthening lymphocyte function-associated antigen 3 on adhesion to CD2.
The effect of lengthening the distance in an adhesion molecule between the receptor binding site and the membrane anchor was studied by inserting four Ig-like domains into the two Ig domain lymphocyte function-associated antigen 3 (LFA-3) molecule. The extended molecule expressed in Chinese hamster ovary (CHO) cells bound to CD2 on T lymphocytes 4- to 20-fold more efficiently than the wild-type molecule at 4 degrees C. Treatment of the CHO clones with neuraminidase to remove sialic acid, or with deoxymannojirimycin to reduce the bulk of N-linked glycosylation, showed that adhesion to both the wild-type and the chimeric LFA-3 molecules was under the influence of cell-cell repulsive forces to a similar extent and that these treatments had less effect than lengthening LFA-3. At higher temperatures, such as 22 and 37 degrees C, the efficiency of binding to the wild-type LFA-3 increased to levels comparable with binding to extended LFA-3. Our results suggest that more distal locations of the adhesive binding site from the cell membrane anchor increase the efficiency of cell-cell adhesion by enhancing the frequency of receptor encounter with ligand and that more proximal locations of the adhesive binding site can provide efficient cell-cell adhesion at physiological temperatures.
Harvard Medical School, Boston, Massachusetts.
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Chan
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Springer
D017485:1-Deoxynojirimycin; D000818:Animals; D000945:Antigens, Differentiation, T-Lymphocyte / Q000235:genetics / Q000378:metabolism*; D018801:CD2 Antigens; D016466:CHO Cells; D050260:Carbohydrate Metabolism; D002241:Carbohydrates / Q000235:genetics; D015815:Cell Adhesion Molecules / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D006224:Cricetinae; D005944:Glucosamine / Q000031:analogs & derivatives / Q000494:pharmacology; D018799:Intercellular Adhesion Molecule-1; D016169:Lymphocyte Function-Associated Antigen-1 / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D009439:Neuraminidase / Q000494:pharmacology; D011971:Receptors, Immunologic / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins; D013329:Structure-Activity Relationship
D000945:Antigens, Differentiation, T-Lymphocyte; D018801:CD2 Antigens; D002241:Carbohydrates; D015815:Cell Adhesion Molecules; D016169:Lymphocyte Function-Associated Antigen-1; D011971:Receptors, Immunologic; D011993:Recombinant Fusion Proteins; D018799:Intercellular Adhesion Molecule-1; D017485:1-Deoxynojirimycin; D009439:Neuraminidase; D005944:Glucosamine