Adhesion of human epidermal keratinocytes to laminin.
We have examined the mechanism by which human epidermal keratinocytes adhere to the A/B1/B2 (alpha 1 beta 1 gamma 1) form of laminin. Adhesion could be completely inhibited with an antibody to the beta 1 integrin subunit or a combination of antibodies recognising the alpha 2 beta 1, alpha 3 beta 1 and alpha 6 beta 4 integrins. Keratinocytes adhered in the presence of magnesium and manganese ions, but calcium ions did not support adhesion and inhibited adhesion when combined with magnesium and manganese. The effects of anti-integrin antibodies (including a stimulatory antibody to the beta 1 subunit) were not influenced by specific cations, with the exception that inhibition by an antibody to alpha 2 beta 1 was abrogated by the presence of manganese ions. The E3 and E8 proteolytic fragments of laminin did not support keratinocyte adhesion and heat inactivation of the E8 site in intact laminin did not reduce adhesion. Three laminin fragments that did support adhesion were P1, E4 and E1X-Nd, P1 activity being attributable at least in part to the RGD site; antibody blocking experiments suggested that adhesion to these fragments was primarily via alpha 3 beta 1. The synthetic peptide GD-6, derived from the carboxy terminus of the laminin A chain (included within E3) did support adhesion, but the significance of this observation is unclear, since a scrambled control peptide could also support adhesion. In conclusion, keratinocyte adhesion to A/B1/B2 laminin involves three integrins and multiple binding sites that are different from those defined previously.
Karecla
P I
PI
Keratinocyte Laboratory, Imperial Cancer Research Fund, London, England.
Timpl
R
R
Watt
F M
FM
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D001667:Binding, Competitive; D002413:Cations, Divalent / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D006801:Humans; D016023:Integrins / Q000502:physiology; D015603:Keratinocytes / Q000166:cytology*; D007797:Laminin / Q000378:metabolism*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D010455:Peptides / Q000138:chemical synthesis / Q000378:metabolism
D000911:Antibodies, Monoclonal; D002413:Cations, Divalent; D016023:Integrins; D007797:Laminin; D009842:Oligopeptides; D010446:Peptide Fragments; D010455:Peptides; C047981:arginyl-glycyl-aspartic acid