Atomic force microscopy-based detection of binding and cleavage site of matrix metalloproteinase on individual type II collagen helices

Hui Bin Sun, Gerald N. Smith, Karen A. Hasty, Hiroki Yokota

Research output: Contribution to journalArticle

33 Scopus citations


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 languageEnglish (US)
Pages (from-to)153-158
Number of pages6
JournalAnalytical Biochemistry
Issue number2
StatePublished - Aug 1 2000
Externally publishedYes



  • Atomic force microscopy
  • MMP-8
  • Matrix metalloproteinase
  • Type II collagen

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this