Dev, Kumlesh K., Chatterjee, Sandipan, Osinde, Maribel, Stauffer, Daniela, Morgan, Hannah, Kobialko, Monika, Dengler, Uwe, Rueeger, Heinrich, Martoglio, Bruno and Rovelli, Giorgio (2006) Signal peptide peptidase dependent cleavage of type II transmembrane substrates releases intracellular and extracellular signals. European Journal of Pharmacology, 540 (1-3). pp. 10-17. ISSN 0014-2999

Abstract

The intramembrane-cleaving proteases (I-CLiPs) presenilin-1 and -2 (PS1 and PS2), signal peptide peptidase (SPP) and the Site-2 protease (S2P) catalyze critical steps in cell signaling and are implicated in diseases such as Alzheimer's disease, hepatitis C virus (HCV) infection and cholesterol homeostasis. Here we describe the development of a cellular assay based on cleavage of the transmembrane sequence of the HCV core protein precursor, releasing intra- and extra-cellular signals that represent sequential signal peptidase and SPP cleavage, respectively. We find that the SPP inhibitor (Z-LL)2-ketone (IC50 = 1.33 microM) and the gamma-secretase potent inhibitors NVP-AHW700-NX (IC50 = 51 nM) and LY411575 (IC50 = 61 nM) but not DAPT dose dependently inhibited SPP but not signal peptidase cleavage. Our data confirm that type II orientated substrates, like the HCV transmembrane sequence, are sequentially cleaved by signal peptidase then SPP. This dual assay provides a powerful tool to pharmacologically analyze sequential cleavage events of signal peptidase and SPP and their regulation.

Item Type:	Article
Related URLs:	http://www.ncbi.nlm.nih.gov/sites/entrez... doi:10.1016/j.ejphar.2006.04.011
Additional Information:	author can archive post-print (ie final draft post-refereeing); Publisher's version/PDF cannot be used
Keywords:	I-CLiP (intramembrane cleaving protease); γ-Secretase inhibitor; SPP (single peptide peptidase) splice variant; S2P (Site-2 protease) cleavage; Reporter gene assay
Related URLs:	http://www.ncbi.nlm.nih.gov/sites/entrez... doi:10.1016/j.ejphar.2006.04.011
Date Deposited:	14 Dec 2009 14:00
Last Modified:	31 Jan 2013 01:17
URI:	https://oak.novartis.com/id/eprint/409