Related Experiment Videos
Hyaluronate binding properties of versican.
R G LeBaron1, D R Zimmermann, E Ruoslahti
1La Jolla Cancer Research Foundation, California 92037.
The Journal of Biological Chemistry
|May 15, 1992
Summary
We found that the large chondroitin sulfate proteoglycan, versican, binds to hyaluronate. This binding activity is located at the N-terminus of versican, demonstrating its role in hyaluronan interactions.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Versican is a large chondroitin sulfate proteoglycan cloned from human fibroblasts.
- Sequence homology at the N-terminus suggests versican may bind hyaluronate.
Purpose of the Study:
- To investigate the hypothesis that versican binds hyaluronate.
- To determine the location of the hyaluronate-binding activity within the versican molecule.
Main Methods:
- Reconstruction and transfection of full-length and N-terminal versican cDNA into mammalian cells (CHO and mouse 3T3 fibroblasts).
- Enzymatic and immunologic analysis of recombinant versican.
- Hyaluronate affinity chromatography to assess binding to hyaluronate, heparin, and chondroitin sulfate.
- Characterization of binding kinetics and determination of dissociation constant.
Main Results:
- Transfected cells produced functional versican and a truncated N-terminal fragment.
- Recombinant versican and its N-terminal fragment specifically bind hyaluronate.
- Binding is concentration-dependent, time-dependent, and can be competed by unlabeled versican.
- Dissociation constant for versican-hyaluronate binding is 4 x 10^-9 M.
Conclusions:
- Versican possesses a specific hyaluronate-binding activity.
- The hyaluronate-binding domain is located at the N-terminus of versican.
- Versican interacts with hyaluronate with high affinity.