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Inhibition of cell adhesion by proteolytic fragments of type V collagen
1Biology Division, National Cancer Center Research Institute, Tokyo, Japan.
Abstract:
Type V collagen inhibits the cell-substratum adhesion of many types of cells. In this study, inhibitory effects of type V collagen on the adhesion of mouse melanoma B16-F10 cells to fibronectin, laminin and vitronectin were investigated. When the culture dishes were coated with a mixture of fibronectin and type V collagen, adhesion of the cells was inhibited by 50% at a fibronectin/collagen molar ratio of 10/1. At a similar molar ratio, adhesion of the cells to laminin was inhibited moderately, but that to vitronectin was not significantly affected. Type V collagen added into culture medium was less effective in inhibiting cell adhesion. The antiadhesive activity of type V collagen was partially retained in the alpha 1 (V) chain of heat-denatured collagen. The alpha 1 (V) chain was split into two large fragments, 90 kDa and 60 kDa, by limited digestion with Staphylococcus aureus V8 proteinase. The 90-kDa fragment, which was derived from the C-terminal half of the alpha 1 (V) chain, inhibited the cell adhesion more profoundly than alpha 1 (V). However, little fibronectin bound to the 90-kDa fragment, while fibronectin bound to the 60-kDa fragment, which was less antiadhesive than the 90-kDa fragment, with the same extent as alpha 1 (V). We therefore concluded that the antiadhesive effect of type V collagen was not due to its specific binding to the fibronectin molecule.
Insights
Type V collagen inhibits melanoma cell adhesion to fibronectin and laminin. This antiadhesive effect is mediated by a specific fragment of the alpha 1 (V) chain, not by direct binding to fibronectin.
Area of Science:
- Biochemistry
- Cell Biology
- Biomaterials Science
Background:
- Type V collagen is known to inhibit cell-substratum adhesion.
- Understanding collagen's role in cell adhesion is crucial for biomaterial design and cancer research.
Purpose of the Study:
- To investigate the inhibitory effects of type V collagen on mouse melanoma B16-F10 cell adhesion.
- To identify the specific regions of type V collagen responsible for its antiadhesive properties.
Main Methods:
- Coating culture dishes with mixtures of fibronectin, laminin, or vitronectin and type V collagen.
- Assessing melanoma cell adhesion rates.
- Analyzing heat-denatured type V collagen and its fragments (alpha 1 (V) chain, 90 kDa, and 60 kDa fragments) for antiadhesive activity.
- Investigating fibronectin binding to collagen fragments.
Main Results:
- Type V collagen significantly inhibited B16-F10 cell adhesion to fibronectin and, to a lesser extent, to laminin, but not to vitronectin.
- The alpha 1 (V) chain retained partial antiadhesive activity.
- A 90-kDa fragment of the alpha 1 (V) chain showed stronger inhibition than the intact chain.
- Fibronectin binding was minimal to the 90-kDa fragment but significant to the 60-kDa fragment.
Conclusions:
- The antiadhesive effect of type V collagen on melanoma cell adhesion is not mediated by direct binding to fibronectin.
- Specific fragments of the alpha 1 (V) chain are responsible for the antiadhesive properties of type V collagen.