Immobilized amines and basic amino acids as mimetic heparin-binding domains for cell surface proteoglycan-mediated adhesion

Stephen Massia, Jeffrey A. Hubbell

Research output: Contribution to journalArticle

83 Citations (Scopus)

Abstract

Diamines covalently coupled to glass substrates promoted human foreskin fibroblast adhesion in the absence of serum. These diamine-derivatized substrates were produced by coupling ethylene diamine, N-methylaminoethylamine, and N,N-dimethylaminoethylamine (NNDMAEA), to sulfonyl chloride-activated glass. Electron spectroscopy for chemical analysis demonstrated that the diamines were coupled via their primary amine ends to produce a surface-bound secondary amine linked to a free amino moiety via a two-carbon spacer. NNDMAEA-modified substrates containing free tertiary amines supported the highest degree of cell spreading (73 ± 7% actively spreading cells) and the most extensive cytoskeletal organization. Both the free tertiary and surface-bound secondary amines were shown to be required for cell spreading. Lysine- and arginine-grafted substrates supported cell spreading and cytoskeletal organization similar to that on NNDMAEA-modified substrates. Although some stress fibers were observed within spread cells on these substrates, focal contacts did not form. Heparinase treatment did not inhibit cell attachment or spreading to the diamine-derivatized substrates, however chondroitinase ABC inhibited cell attachment and spreading on all substrates; heparinase inhibited spreading on lysine- and arginine-derivatized substrates to a lesser extent. These results imply that cell attachment to these substrates was mediated primarily by cell surface chondroitin sulfate proteoglycans. This study demonstrates that covalently grafted NNDMAEA, lysine, and arginine can mimic the adhesion-promoting activity of the glycosaminoglycan-binding domains of cell adhesion proteins. This study also demonstrates that the interaction with these proteoglycans depends in a very sensitive manner on the particular structure of the immobilized amine.

Original languageEnglish (US)
Pages (from-to)10133-10141
Number of pages9
JournalJournal of Biological Chemistry
Volume267
Issue number14
StatePublished - May 15 1992
Externally publishedYes

Fingerprint

Basic Amino Acids
Proteoglycans
Amines
Heparin
Adhesion
Diamines
Substrates
Heparin Lyase
Lysine
Arginine
Glass
Chondroitin ABC Lyase
Chondroitin Sulfate Proteoglycans
Foreskin
Photoelectron Spectroscopy
Stress Fibers
Focal Adhesions
Electron spectroscopy
Glycosaminoglycans
Cell adhesion

ASJC Scopus subject areas

  • Biochemistry

Cite this

Immobilized amines and basic amino acids as mimetic heparin-binding domains for cell surface proteoglycan-mediated adhesion. / Massia, Stephen; Hubbell, Jeffrey A.

In: Journal of Biological Chemistry, Vol. 267, No. 14, 15.05.1992, p. 10133-10141.

Research output: Contribution to journalArticle

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