TopFIND 4.0

Evidence Source

Description Inferred from cleavage recMMP-Chemokine_AStarr2012A: 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.

Method

electronic annotation

Experimental system

unknown

Perturbation

none

Protease inhibitor(s) used


Directness

indirect

Physiological relevance

unknown

Evidence Code(s)

    inferred from cleavage

Database / Laboratory

Raw data repository

Evidence for:

1

N-termini

1

C-termini


Publication(s)