Synthetic peptides from the carboxy-terminal globular domain of the A chain of laminin: their ability to promote cell adhesion and neurite outgrowth, and interact with heparin and the beta 1 integrin subunit.
Open Access
- 15 November 1991
- journal article
- Published by Rockefeller University Press in The Journal of cell biology
- Vol. 115 (4), 1137-1148
- https://doi.org/10.1083/jcb.115.4.1137
Abstract
The large carboxy-terminal globular domain (G domain; residues 2,110-3,060) of the A chain of murine-derived laminin has been shown to promote heparin binding, cell adhesion, and neurite outgrowth. This study was conducted to define the potential sequence(s) originating from the G domain of laminin with any of these functional activities. A series of peptides were synthesized from the G domain, termed GD peptides, each approximately 20 amino acids long and containing multiple positively charged amino acids. In direct 3H-heparin binding assays, peptides GD-1 and GD-2 bound high levels of 3H-heparin, while peptides GD-3 and GD-4 bound lower levels of 3H-heparin, and GD-5 bound essentially no 3H-heparin. The binding of 3H-heparin to peptides GD-1 and GD-2 appeared to be of high affinity, since significant binding of 3H-heparin to these two peptides was still observed even when the NaCl concentration was raised to 1.0 M. Four of the peptides, GD-1, GD-2, GD-3, and GD-4, directly promoted the adhesion and spreading of HT-1080 human fibrosarcoma cells as well as the outgrowth of neurites from chick spinal cord and dorsal root ganglia neurons. In addition, solutions of these peptides or antibodies generated against these peptides inhibited laminin-mediated HT-1080 cell adhesion. Antibodies against the beta 1 integrin subunit inhibited HT-1080 cell adhesion and neurite outgrowth on surfaces adsorbed with peptides GD-3 and GD-4. Therefore, laminin appears to have multiple, independent sequences in the G domain that serve a similar cell adhesion promoting function for different cell types. Furthermore, these results suggest that the sequences comprising peptides GD-3 and GD-4 use an integrin as a receptor, of which the beta 1 integrin subunit is a component for these various cell types.Keywords
This publication has 61 references indexed in Scilit:
- A neuronal cell line (PC12) expresses two β1-class integrins—α1β1, and α3β1—that recognize different neurite outgrowth-promoting domains in lamininNeuron, 1990
- Cell adhesion, spreading and neurite stimulation by laminin fragment E8 depends on maintenance of secondary and tertiary structure in its rod and globular domainEuropean Journal of Biochemistry, 1990
- The role of integrins alpha 2 beta 1 and alpha 3 beta 1 in cell-cell and cell-substrate adhesion of human epidermal cells.The Journal of cell biology, 1990
- Laminin a chain synthetic peptide which supports neurite outgrowthBiochemical and Biophysical Research Communications, 1989
- Purification and characterization of mammalian integrins expressed by a rat neuronal cell line (PC12): evidence that they function as alpha/beta heterodimeric receptors for laminin and type IV collagen.The Journal of cell biology, 1988
- N-cadherin and integrins: Two receptor systems that mediate neuronal process outgrowth on astrocyte surfacesNeuron, 1988
- Localization of a tumor cell adhesion domain of laminin by a monoclonal antibodyExperimental Cell Research, 1987
- Interactions of a neuronal cell line (PC12) with laminin, collagen IV, and fibronectin: identification of integrin-related glycoproteins involved in attachment and process outgrowth.The Journal of cell biology, 1987
- Mapping of domains in human laminin using monoclonal antibodies: localization of the neurite-promoting site.The Journal of cell biology, 1986
- Protease Resistance and Conformation of LamininEuropean Journal of Biochemistry, 1982