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 Hasty, Hiroki Yokota

Research output: Contribution to journalArticle

33 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)153-158
Number of pages6
JournalAnalytical Biochemistry
Volume283
Issue number2
DOIs
StatePublished - Aug 1 2000

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Matrix Metalloproteinase 8
Collagen Type II
Atomic Force Microscopy
Matrix Metalloproteinases
Atomic force microscopy
Collagen
Binding Sites
Tropocollagen
Molecules
Collagenases
Visualization
Enzymes
Proteins
Substrates

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Atomic force microscopy-based detection of binding and cleavage site of matrix metalloproteinase on individual type II collagen helices. / Sun, Hui Bin; Smith, Gerald N.; Hasty, Karen; Yokota, Hiroki.

In: Analytical Biochemistry, Vol. 283, No. 2, 01.08.2000, p. 153-158.

Research output: Contribution to journalArticle

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