Collagen targeting using multivalent protein-functionalized dendrimers.
Collagen is an attractive marker for tissue remodeling in a variety of common disease processes. Here we report the preparation of protein dendrimers as multivalent collagen targeting ligands by native chemical ligation of the collagen binding protein CNA35 to cysteine-functionalized dendritic divalent (AB(2)) and tetravalent (AB(4)) wedges. The binding of these multivalent protein constructs was studied on collagen-immobilized chip surfaces as well as to native collagen in rat intestinal tissues. To understand the importance of target density we also created collagen-mimicking surfaces by immobilizing synthetic collagen triple helical peptides at various densities on a chip surface. Multivalent display of a weak-binding variant (CNA35-Y175K) resulted in a large increase in collagen affinity, effectively restoring the collagen imaging capacities for the AB(4) system. In addition, dissociation of these multivalent CNA35 dendrimers from collagen surfaces was found to be strongly attenuated.
Breurken
Monica
M
Laboratory of Chemical Biology, Department of Biomedical Engineering, Eindhoven University of Technology, PO Box 513, 5600 MB Eindhoven, The Netherlands.
Lempens
Edith H M
EH
Temming
Rinske P
RP
Helms
Brett A
BA
Meijer
E W
EW
Merkx
Maarten
M
D000818:Animals; D003094:Collagen / Q000378:metabolism*; D003545:Cysteine / Q000378:metabolism; D050091:Dendrimers / Q000138:chemical synthesis / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D008024:Ligands; D057054:Molecular Imaging; D010957:Plasmids; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D020349:Surface Plasmon Resonance / Q000379:methods*
D050091:Dendrimers; D008024:Ligands; D011506:Proteins; D003094:Collagen; D003545:Cysteine