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Covalent Surface Immobilization of Arg-Gly-Asp- And Tyr-Ile-Gly-Ser-Arg-containing Peptides to Obtain Well-Defined Cell-Adhesive Substrates

Covalent Surface Immobilization of Arg-Gly-Asp- And Tyr-Ile-Gly-Ser-Arg-containing Peptides to Obtain Well-Defined Cell-Adhesive Substrates

S P Massia, J A Hubbell

Anal Biochem. 1990 Jun;187(2):292-301.

PMID: 2382830

Abstract:

The synthetic peptides Gly-Arg-Gly-Asp-Tyr and Gly-Tyr-Ile-Gly-Ser-Arg-Tyr, which contain Arg-Gly-Asp (RGD) and Tyr-Ile-Gly-Ser-Arg (YIGSR), the ligands for two important classes of cell adhesion receptors, were covalently coupled to a nonadhesive modified glass surface by the N-terminal Gly. The N-terminal Gly served as a spacer, and the C-terminal Y served as a site for radioiodination. These modified substrates supported the adhesion and spreading of cultured human foreskin fibroblasts (HFFs) independently of adsorbed proteins and, it was demonstrated that a covalently immobilized YIGSR-containing peptide has biological activity. The surface concentration of grafted peptide on the glass was measured by 125I radio-labeling and was 12.1 pmol/cm2. HFFs spread on both immobilized peptide substrates, but at much slower rates on grafted YIGSR glass surfaces than on the RGD-containing substrates. Cells formed focal contacts on the RGD-derivatized substrates in the presence or absence of serum. Focal contacts formed on the YIGSR-grafted surfaces only when serum was present in the medium and had morphologies different from those observed on the RGD-containing substrates. Serum influenced the organization of microfilaments and the extent of spreading of adherent cells, although adsorption of adhesion proteins was minimal on all substrates. This derivatization method produced chemically stable substrates which may be useful in studying receptor-mediated cell adhesion, as the quantity of peptide available at the surface may be precisely measured and controlled.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP110590642	Tyr-Ile-Gly-Ser-Arg		110590-64-2	Price

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