Atomic Force Microscopy-Based Detection of Binding and Cleavage Site of Matrix Metalloproteinase on Individual Type II Collagen Helices
✍ Scribed by Hui Bin Sun; Gerald N. Smith Jr.; Karen A. Hasty; Hiroki Yokota
- Publisher
- Elsevier Science
- Year
- 2000
- Tongue
- English
- Weight
- 222 KB
- Volume
- 283
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
Type II tropocollagen molecules were reacted with matrix metalloproteinase 8 (MMP-8) and the binding sites as well as the cleavage site of MMP-8 were detected on individual molecules using atomic force microscopy (AFM). Approximately 300-nm-long coiledcoil tropocollagen molecules were straightened and immobilized on an atomically flat surface for detection by AFM. The direct visualization of individual collagen molecules revealed heterogeneous characteristics of MMP-8:collagen complexes. We observed that there existed multiple MMP-8 nonspecific binding sites on the collagen molecules, but cleavage always took place at a unique site. When collagen molecules, straightened and immobilized on the surface, were reacted with MMP-8, a site of cleavage appeared as a gap in stretched molecules. This is the first report to visually show direct collagenase:collagen interactions using AFM. The described AFM-based analysis has potential as a protein analysis tool for understanding a complex mechanism of enzyme:substrate interactions.