Photoimmobilization of a bioactive laminin fragment and pattern-guided selective neuronal cell attachment.
To attain light-dependent functionalization of biocompatible materials, a photolabel-derivatized, bioactive laminin fragment has been synthesized, chemically characterized, and photoimmobilized. Covalent high-resolution patterning of the laminin fragment CDPGYIGSR to hydroxylated fluorinated ethylene propylene (FEP-OH), poly(vinyl alcohol), and glycophase glass has been achieved. The synthetic peptide CDPGYIGSR was thermochemically coupled to either N-[m-[3-(trifluoromethyl)-diazirin-3-yl]phenyl]-4-maleimidobuty ramide or 4-maleimidobenzophenone. Photolabel-derivatized peptides were radiolabeled, and 20 and 300 microns-sized patterns were visualized by autoradiography. The biospecific interaction of photoimmobilized laminin fragments with cells was investigated by analyzing the selective attachment of NG 108-15 neuroblastoma x glioma cells which bear CDPGYIGSR-specific cell surface receptors. On photopatterned FEP-OH membranes NG 108-15 cells differentiated in serum-supplemented media within 1 day. Specific attachment to the immobilized oligopeptide CDPGYIGSR was assessed in serum-free media with competitive binding studies, showing an 82% decrease in cell adherence after the cell receptors were blocked with soluble CDPGYIGSR.
Clémence
J F
JF
Institute of Biochemistry, University of Bern, Switzerland.
Ranieri
J P
JP
Aebischer
P
P
Sigrist
H
H
D000595:Amino Acid Sequence; D000818:Animals; D001389:Azirines; D001577:Benzophenones; D001667:Binding, Competitive; D002448:Cell Adhesion; D018914:Cells, Immobilized; D005898:Glass; D005910:Glioma; D006822:Hybrid Cells; D007202:Indicators and Reagents; D007797:Laminin; D008301:Maleimides; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009447:Neuroblastoma; D009474:Neurons / Q000166:cytology* / Q000502:physiology*; D010446:Peptide Fragments / Q000138:chemical synthesis; D010777:Photochemistry; D011138:Polytetrafluoroethylene / Q000031:analogs & derivatives; D011142:Polyvinyl Alcohol; D011956:Receptors, Cell Surface / Q000502:physiology; D005465:Fluorocarbon Polymers
D001389:Azirines; D001577:Benzophenones; D007202:Indicators and Reagents; D007797:Laminin; D008301:Maleimides; D010446:Peptide Fragments; D011138:Polytetrafluoroethylene; D011142:Polyvinyl Alcohol; D011956:Receptors, Cell Surface; C096305:fluorinated ethylene propylene; D005465:Fluorocarbon Polymers; C085785:N-(2-(3-(trifluoromethyl)diazirin-3-yl)phenyl)-4-maleimidobutyramide; C046109:benzophenone-4-maleimide