PMID: 9546670Apr 18, 1998Paper

One-step purification of cathepsin D by affinity chromatography using immobilized propeptide sequences

European Journal of Biochemistry
S WittlinD R Stover

Abstract

In vivo, active cathepsin D proteinase is generated by removal of a 44-residue propeptide at its N-terminus. Here we report that mature cathepsin D and pseudocathepsin D (a partially activated form of cathepsin D with 25 amino acid residues removed from the propeptide) bind to the immobilized propeptide, while procathepsin D does not. The N-terminal 25 amino acid residues of the propeptide are sufficient for this binding. Based on this observation, a simple one-step procedure was developed to purify mature cathepsin D from whole cell extracts to near homogeneity. This method has the advantage over existing affinity-purification systems that active forms of the proteinase can be separated from inactive precursors and other aspartic proteinases. Furthermore, this technique was effective for pepsin as well, suggesting it may have general utility for all activated aspartic proteinases and perhaps other families of proteinases.

Citations

Dec 14, 2004·Chembiochem : a European Journal of Chemical Biology·Norbert SchaschkeChristian P Sommerhoff
Nov 6, 2001·Journal of Biochemical and Biophysical Methods·R Zusman, I Zusman
Feb 24, 2001·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·L B Nielsen, H H Nielsen
Oct 23, 1998·Nature Structural Biology·A Y LeeJ W Erickson
Apr 6, 2000·The Biochemical Journal·S J SandersonG H Coombs
Sep 22, 1999·European Journal of Biochemistry·S WittlinD R Stover
Apr 28, 2000·European Journal of Biochemistry·V Schauer-VukasinovicT Giller
Apr 6, 1999·APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica·H Rochefort, E Liaudet-Coopman
Feb 26, 2008·Folia Histochemica Et Cytobiologica·Alina MinarowskaŁukasz Minarowski
Dec 14, 2011·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Visa Merino, Nadimpalli Siva Kumar
Apr 13, 2007·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Pål Anders WangRagnar L Olsen
Apr 2, 2002·Protein Expression and Purification·Sergei V GulnikJohn W Erickson
Sep 17, 2010·Journal of Agricultural and Food Chemistry·Rafik BaltiMoncef Nasri

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