A laminin-pepsin fragment with cell attachment and neurite outgrowth activity at distinct sites.
Laminin is a large basement membrane glycoprotein which influences the behavior and morphology of a variety of cells. We have found that laminin and a pepsin fragment of laminin (P-lam) contain distinct sites for HT-1080 human fibrosarcoma cell attachment and for neurite outgrowth activity of PC12 and NG108-15 cell lines. Reduction and alkylation of laminin and P-lam fragment disulfide bonds, in the absence of denaturing agents, markedly reduced the cell attachment activity without reducing the neurite outgrowth response. The P-lam fragment (approximately 375 kDa) was found to contain part of the cross region of laminin and a portion of the long arm, on the basis of recognition by antisera against laminin synthetic peptides and fusion proteins. Modification of arginine residues by cyclohexanedione also had no effect on neurite outgrowth but reduced HT-1080 cell adhesion. Modification of lysine residues by succinic and citraconic anhydride, however, abolished laminin neurite outgrowth but not cell attachment activity. Neurite outgrowth activity was recovered by reversing the lysine modification. These data support the existence on laminin of separate sites for cell attachment and for neurite outgrowth.
Sephel
G C
GC
Laboratory of Developmental Biology and Anomalies National Institute of Dental Research, Bethesda, Maryland 20892.
Tashiro
K
K
Sasaki
M
M
Kandel
S
S
Yamada
Y
Y
Kleinman
H K
HK
D000478:Alkylation; D000818:Animals; D001665:Binding Sites; D001681:Biological Assay; D015153:Blotting, Western; D002448:Cell Adhesion; D002465:Cell Movement; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D006493:Heparin / Q000378:metabolism; D066298:In Vitro Techniques; D007797:Laminin / Q000378:metabolism / Q000648:ultrastructure*; D008970:Molecular Weight; D009474:Neurons / Q000166:cytology*; D010084:Oxidation-Reduction; D010434:Pepsin A; D010446:Peptide Fragments / Q000378:metabolism; D011487:Protein Conformation; D051381:Rats; D013329:Structure-Activity Relationship
D007797:Laminin; D010446:Peptide Fragments; D006493:Heparin; D010434:Pepsin A