Efficient accommodation of recombinant, foot-and-mouth disease virus RGD peptides to cell-surface integrins.
The engineering of either complete virus cell-binding proteins or derived ligand peptides generates promising nonviral vectors for cell targeting and gene therapy. In this work, we have explored the molecular interaction between a recombinant, integrin-binding foot-and-mouth disease virus RGD peptide displayed on the surface of a carrier protein and its receptors on the cell surface. By increasing the number of viral segments, cell binding to recombinant proteins was significantly improved. This fact resulted in a dramatic growth stimulation of virus-sensitive BHK(21) cells but not virus-resistant HeLa cells in protein-coated wells. Surprisingly, growth stimulation was not observed in vitronectin-coated plates, suggesting that integrins other than alpha(v)beta(3) could be involved in binding of the recombinant peptide, maybe as coreceptors. On the other hand, both free and cell-linked integrins did not modify the enzymatic activity of RGD-based enzymatic sensors that contrarily, were activated by the induced fit of anti-RGD antibodies. Those findings are discussed in the context of a proper mimicry of the unusually complex architecture of this cell-binding site as engineered in multifunctional proteins.
Alcalá
P
P
Institut de Biotecnologia i de Biomedicina, Departament de Genètica i de Microbiologia, Universitat Autònoma de Barcelona, Bellaterra, 08193 Barcelona, Spain.
Feliu
J X
JX
Arís
A
A
Villaverde
A
A
D000595:Amino Acid Sequence; D000818:Animals; D005537:Aphthovirus / Q000235:genetics / Q000502:physiology*; D002455:Cell Division; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology* / Q000821:virology; D006224:Cricetinae; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015316:Genetic Therapy; D006367:HeLa Cells; D006801:Humans; D016023:Integrins / Q000502:physiology*; D007700:Kinetics; D008958:Models, Molecular; D009842:Oligopeptides; D011487:Protein Conformation; D017469:Receptors, Fibronectin / Q000502:physiology; D011991:Receptors, Virus / Q000502:physiology; D019030:Receptors, Vitronectin / Q000502:physiology; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D011995:Recombination, Genetic; D019096:Vitronectin / Q000502:physiology; D001616:beta-Galactosidase / Q000737:chemistry / Q000378:metabolism
D016023:Integrins; D009842:Oligopeptides; D017469:Receptors, Fibronectin; D011991:Receptors, Virus; D019030:Receptors, Vitronectin; D011994:Recombinant Proteins; D019096:Vitronectin; C047981:arginyl-glycyl-aspartic acid; D001616:beta-Galactosidase