Accessible Requires Authentication Published by De Gruyter June 18, 2011

Outside or inside: role of the subcellular localization of DP4-like enzymes for substrate conversion and inhibitor effects

Ute Bank, Anke Heimburg, Astrid Wohlfarth, Gudrun Koch, Karsten Nordhoff, Heiko Julius, Martin Helmuth, Doreen Breyer, Dirk Reinhold, Michael Täger and Siegfried Ansorge
From the journal

Abstract

The discovery of the DP4-related enzymes DP8 and DP9 raised controversial discussion regarding the physiological and pathophysiological function of distinct members of the DP4 family. Particularly with regard to their potential relevance in regulating immune functions, it is of interest to know which role the subcellular distribution of the enzymes play. Synthetic substrates as well as low molecular weight inhibitors are widely used as tools, but little is yet known regarding their features in cell experiments, such as their plasma membrane penetration capacity. The fluorogenic substrates Gly-Pro-AMC or (Ala-Pro)2-R110 predominantly detect plasma membrane-bound activities of viable cells (less than 0.1% of fluorochromes R110 or AMC inside viable cells after 1 h incubation). Additionally, the selective and non-selective DP8/9 inhibitors allo-Ile-isoindoline and Lys[Z(NO2)]-pyrrolidide were found to be incapable of passing the plasma membrane easily. This suggests that previously reported cellular effects are not due to inhibition of the cytosolic enzymes DP8 or DP9. Moreover, our enzymatic studies with viable cells provided evidence that DP8 and/or DP9 are also present on the surface of immune cells under certain circumstances and could gain relevance particularly in the absence of DP4 expression. In summary, in cells which do express DP4 on the surface, this archetypical member of the DP4 family is the most relevant peptidase in the regulation of cellular functions.


Corresponding author

Received: 2010-7-5
Accepted: 2010-12-1
Published Online: 2011-06-18
Published in Print: 2011-03-01

©2011 by Walter de Gruyter Berlin New York