TopFIND 4.0

Evidence Source

Description Inferred from cleavage recMMP-Chemokine_AStarr2012B: Cleavage Assays—In vitro native substrate cleavage assays performed at enzyme to substrate ratios of 1:10 (w/w) of chemokines, vimentin, IGFBP-7, cystatin C, galectin-1, and secreted protein, acidic and rich in cysteine (SPARC) by sMT6- MMPdeltaF were performed in a 10-µl reaction containing 50 mM HEPES, 200 mM NaCl, 5 mM CaCl2, pH 7.4 for 16 h at 37 °C. Chemokine cleavage assay products were analyzed by MALDI-TOF mass spectrometry (MS) on a Voyager-DE STR (Applied Biosystems) using sinapinic acid matrix (33) and confirmed by silver-stained 15% Tris-Tricine SDS-PAGE. Chemokine cleavage was defined to be positive when the mass spectrometry spectra showed a cleavage product with ~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)