Evidence Source
| Description | In vitro chemokine cleavage assays with recombinant MMPs were performed in 50 mM Tris, 200 mM NaCl, 5 mM CaCl2, pH 7.4 for 16 h at 37 °C. Initially, an enzyme to substrate molar ratio of 1:10 was assessed to screen for cleavage activity, followed by lower enzyme concentrations to 1:1,000 to suggest biological relevance. Cleavage assay products were analyzed, as above, by MALDI-TOF MS and confirmed by silver-staining following 15% Tris-Tricine SDS- PAGE. Chemokine cleavage was defined to be positive when the MS spectra showed a cleavage product with greater than 20% ion intensity of the full-length chemokine. |
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Method |
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Experimental system |
cell free |
Perturbation |
none |
Protease inhibitor(s) used |
|
Directness |
direct |
Physiological relevance |
unknown |
Evidence Code(s) |
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Database / Laboratory |
- Lab: Overall Lab |
Raw data repository |
Evidence for:
0
N-termini
0
C-termini
Publication(s) |
Biochemical analysis of matrix metalloproteinase activation of chemokines CCL15 and CCL23 and increased glycosaminoglycan binding of CCL16. Starr AE, Dufour A, Maier J, Overall CM J Biol Chem. 2012 Feb 17;287(8):5848-60. doi: 10.1074/jbc.M111.314609. Epub 2011 Dec 6. PMID: 22147696 |
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