A single step purification for autolytic zinc proteinases

Erin M Wilfong; Ursala Locklear; Eric J Toone

doi:10.1016/j.bmcl.2009.10.114

A single step purification for autolytic zinc proteinases

Bioorg Med Chem Lett. 2010 Jan 1;20(1):280-2. doi: 10.1016/j.bmcl.2009.10.114. Epub 2009 Nov 5.

Authors

Erin M Wilfong¹, Ursala Locklear, Eric J Toone

Affiliation

¹ Department of Chemistry, Duke University, LSRC B120, Durham, NC 27708, United States.

Abstract

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.

MeSH terms

Autolysis
Catalytic Domain
Chromatography, Affinity
Histidine / chemistry
Imidazoles / chemistry
Matrix Metalloproteinase 3 / isolation & purification*
Oligopeptides / chemistry
Protein Folding

Substances

His-His-His-His-His-His
Imidazoles
Oligopeptides
Histidine
Matrix Metalloproteinase 3
imidazoleacetic acid

Grants and funding

R01 GM057179/GM/NIGMS NIH HHS/United States