Abstract
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. (C) 2000 Academic Press.
Original language | English (US) |
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Pages (from-to) | 153-158 |
Number of pages | 6 |
Journal | Analytical Biochemistry |
Volume | 283 |
Issue number | 2 |
DOIs | |
State | Published - Aug 1 2000 |
Externally published | Yes |
Keywords
- Atomic force microscopy
- MMP-8
- Matrix metalloproteinase
- Type II collagen
ASJC Scopus subject areas
- Biophysics
- Biochemistry
- Molecular Biology
- Cell Biology