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Purification and characterization of native type XIV collagen
E Aubert-Foucher1, B Font, D Eichenberger
1Institute of Biology and Chemistry of Proteins (CNRS-UPR 412), Lyon, France.
The Journal of Biological Chemistry
|August 5, 1992
Summary
Researchers purified and characterized type XIV collagen, a newly discovered molecule. This study details its cross-shaped structure and molecular properties, differentiating it from type XII collagen.
Area of Science:
- Biochemistry
- Molecular Biology
- Extracellular Matrix Research
Background:
- Type XIV collagen, homologous to types IX and XII, was recently identified in fetal bovine tissues.
- Characterization of its intact native form was previously lacking.
Purpose of the Study:
- To purify and characterize the intact native form of type XIV collagen.
- To compare type XIV collagen with type XII collagen.
Main Methods:
- Two-step chromatographic purification of type XIV collagen.
- Simultaneous large-scale purification of types XII and XIV collagens.
- Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions.
- Collagenase treatment and rotary shadowing electron microscopy.
- Analysis of molecular mass and cross-linking of collagen chains.
Main Results:
- Pure type XIV collagen was obtained using two chromatographic steps.
- Intact type XIV collagen exhibits a cross-shaped structure with a tail, central globule, and three 'fingers'.
- SDS-PAGE revealed bands at 220 and 290 kDa, reducing to 190 kDa after collagenase treatment.
- Type XIV collagen chains (approx. 220 kDa) are linked by disulfide bridges, not other cross-links.
- Differences in charge and glycosylation distinguish type XIV from type XII collagen.
Conclusions:
- Type XIV collagen has been successfully purified and characterized.
- Its structure and properties are distinct from type XII collagen, despite homologies.
- The study provides a foundation for understanding type XIV collagen's biological role.