A single step purification for autolytic zinc proteinases.
We describe a novel single-step method for the purification of stromelysin-1 catalytic domain (SCD) via immobilized metal affinity chromatography under denaturing conditions that inhibit proteolytic activity followed by on-column refolding and spontaneous autolysis of the fusion peptide to yield pure, active stromelysin-1 catalytic domain. The methodology provides a general approach for the rapid purification of large quantities of zinc proteinases.
Wilfong
Erin M
EM
Department of Chemistry, Duke University, LSRC B120, Durham, NC 27708, United States.
Locklear
Ursala
U
Toone
Eric J
EJ
D001329:Autolysis; D020134:Catalytic Domain; D002846:Chromatography, Affinity; D006639:Histidine / Q000737:chemistry; D007093:Imidazoles / Q000737:chemistry; D019278:Matrix Metalloproteinase 3 / Q000302:isolation & purification*; D009842:Oligopeptides / Q000737:chemistry; D017510:Protein Folding
C471213:His-His-His-His-His-His; D007093:Imidazoles; D009842:Oligopeptides; D006639:Histidine; D019278:Matrix Metalloproteinase 3; C005954:imidazoleacetic acid