(Arg-Gly-Asp)n-albumin conjugates as a model substratum for integrin-mediated cell adhesion.
We have prepared protein-peptide conjugates composed of bovine serum albumin (BSA) derivatized with short peptides containing the Arg-Gly-Asp (RGD) sequence derived from the adhesion site of fibronectin. The RGD-BSA conjugates were used to coat tissue culture plastic surfaces which then served as substrata in cell adhesion experiments. Our results indicate that the efficiency of adhesion to RGD-BSA-coated surfaces is highly dependent on the valency of the (RGD)n-BSA conjugates. For example, on surfaces with approximately equal amounts of RGD ligand, CHO cells adhered virtually 100% to the (RGD)n-BSA (n = 20.8) conjugate and not at all to the (RGD)n-BSA (n = 3.5) conjugate. Adhesion on (RGD)n-BSA-coated substrata and on fibronectin- or vitronectin-coated substrata was also examined in terms of the relationship between cell adhesion and the intermolecular distances of adsorbed proteins. It was observed that for substrata coated with relatively compact, symmetric molecules, such as RGD-BSA or vitronectin, adhesion dropped off sharply as intermolecular distances increased; by contrast, for fibronectin, a large asymmetric molecule, adhesion declined more gradually as intermolecular distances increased. Finally, we have examined the role of different cell-surface receptors in the process of adhesion to RGD-BSA substrata. Interestingly, competition and blocking experiments with antibodies and with soluble competing proteins suggest that it is the vitronectin receptor rather than the fibronectin receptor which mediates adhesion to RGD-BSA.
Danilov
Y N
YN
Department of Pharmacology, School of Medicine, University of North Carolina, Chapel Hill 27599-7365.
Juliano
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RL
D000327:Adsorption; D000595:Amino Acid Sequence; D000818:Animals; D000954:Antigens, Surface / Q000502:physiology*; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules; D002460:Cell Line; D006224:Cricetinae; D005353:Fibronectins / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D016023:Integrins; D008562:Membrane Glycoproteins / Q000502:physiology*; D010969:Plastics; D017469:Receptors, Fibronectin; D011971:Receptors, Immunologic / Q000502:physiology*; D019030:Receptors, Vitronectin; D012710:Serum Albumin, Bovine; D013329:Structure-Activity Relationship; D019096:Vitronectin
D000954:Antigens, Surface; D015815:Cell Adhesion Molecules; D005353:Fibronectins; D006023:Glycoproteins; D016023:Integrins; D008562:Membrane Glycoproteins; D010969:Plastics; D017469:Receptors, Fibronectin; D011971:Receptors, Immunologic; D019030:Receptors, Vitronectin; D019096:Vitronectin; D012710:Serum Albumin, Bovine