A signal peptide peptidase (SPP) reporter activity assay based on the cleavage of type II membrane protein substrates provides further evidence for an inverted orientation of the SPP active site relative to presenilin.

The Journal of Biological Chemistry
Andrew C NyborgTodd E Golde

Abstract

Signal peptide peptidase (SPP) is an intramembrane-cleaving protease identified by its cleavage of several type II membrane signal peptides after signal peptidase cleavage. Here we describe a novel, quantitative, cell-based SPP reporter assay. This assay utilizes a substrate consisting of the NH2 terminus of the ATF6 transcription factor fused to a transmembrane domain susceptible to SPP cleavage in vitro. In cells, cleavage of the substrate releases ATF6 from the membrane. This cleavage can be monitored by detection of an epitope that is unmasked in the cleaved substrate or by luciferase activity induced by the cleaved ATF6 substrate binding to and activating an ATF6 luciferase reporter construct. Using this assay we show that (i) SPP is the first aspartyl intramembrane-cleaving protease whose activity increases proportionally to its overexpression and (ii) selectivity of various SPP and gamma-secretase inhibitors can be rapidly evaluated. Because this assay was designed based on data suggesting that SPP has an orientation distinct from presenilin and cleaves type II membrane proteins, we determined whether the segment of SPP located between the two presumptive catalytic aspartates was in the lumen or cytoplasm. Using site-dir...Continue Reading

References

Oct 1, 1991·Immunology Today·K KärreR Kiessling
Apr 1, 1973·Virology·F L Graham, A J van der Eb
Jun 17, 2000·The Journal of Biological Chemistry·Y WangR Prywes
May 30, 2001·Trends in Molecular Medicine·T E Golde, S G Younkin
Nov 21, 2001·The Journal of Immunology : Official Journal of the American Association of Immunologists·M K LembergB Martoglio
Apr 30, 2002·Human Molecular Genetics·Chris P PontingTodd E Golde
Jun 22, 2002·Science·Andreas WeihofenBruno Martoglio
Jul 26, 2002·Biochemistry. Biokhimii︠a︡·A P GrigorenkoE I Rogaev
Nov 7, 2002·Molecular Cell·Marius K Lemberg, Bruno Martoglio
Jan 22, 2003·Biochemical and Biophysical Research Communications·Sébastien S HébertGeorges Lévesque
Mar 7, 2003·The Journal of Biological Chemistry·Andreas WeihofenBruno Martoglio
Mar 7, 2003·Science's STKE : Signal Transduction Knowledge Environment·Todd E Golde, Christopher B Eckman
Mar 20, 2003·The Journal of Biological Chemistry·Anna Y KornilovaMichael S Wolfe
Mar 28, 2003·Nature·Nobumasa TakasugiTakeshi Iwatsubo
Apr 8, 2003·Nature Cell Biology·Dieter EdbauerChristian Haass
Apr 15, 2003·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·M Paul MurphyTodd E Golde
May 13, 2003·Proceedings of the National Academy of Sciences of the United States of America·W Taylor KimberlyDennis J Selkoe
May 24, 2003·Biochemical and Biophysical Research Communications·Laura MarlowKumar Sambamurti
Jun 17, 2003·Journal of Cell Science·Weiming Xia, Michael S Wolfe
Oct 21, 2003·Proceedings of the National Academy of Sciences of the United States of America·Eric H SchroeterRaphael Kopan
Dec 4, 2003·Biochemical Society Transactions·B Martoglio
Jan 6, 2004·The Journal of Biological Chemistry·Andrew C NyborgTodd E Golde

❮ Previous
Next ❯

Citations

Feb 2, 2008·Cellular and Molecular Life Sciences : CMLS·A J Beel, C R Sanders
May 13, 2009·Journal of the American Chemical Society·Yuki ImamuraTsunehiko Higuchi
Jun 2, 2005·Nature Reviews. Immunology·John R Rodgers, Richard G Cook
Jun 4, 2011·The Journal of Biological Chemistry·Hiroyuki MiyashitaTakeshi Iwatsubo
Nov 8, 2012·The Journal of Biological Chemistry·Matthias VossRegina Fluhrer
Nov 5, 2008·The Journal of Biological Chemistry·Thalia RobakisPeter Scheiffele
Feb 5, 2009·The Journal of Biological Chemistry·Regina FluhrerChristian Haass
Dec 26, 2012·The Journal of Experimental Medicine·Daniel R BeisnerMichael P Cooke
Aug 19, 2005·Molecular Biology of the Cell·Shirly Becker-HermanIdit Shachar
Jun 13, 2012·Microbiology and Molecular Biology Reviews : MMBR·Ross E DalbeyJan Maarten van Dijl
Jun 29, 2007·Neuro-degenerative Diseases·Regina Fluhrer, Christian Haass
May 1, 2009·Expert Review of Neurotherapeutics·Taisuke Tomita
May 12, 2009·Seminars in Cell & Developmental Biology·Todd E GoldeDoron C Greenbaum
May 30, 2006·Biochimica Et Biophysica Acta·Jens UrnyH Chica Schaller
Feb 22, 2014·Current Opinion in Immunology·Sander I van KasterenJacques Neefjes
Jun 26, 2013·Biochimica Et Biophysica Acta·Todd E GoldeLucio Miele
Oct 9, 2013·Biochimica Et Biophysica Acta·Matthias VossRegina Fluhrer
Jul 16, 2014·Biochemical and Biophysical Research Communications·Michael BaldwinAthar H Chishti
Jun 19, 2017·Biochimica Et Biophysica Acta. Molecular Cell Research·Torben MentrupBernd Schröder
Jul 28, 2005·The Journal of Biological Chemistry·Hanna LaudonJan Näslund
Jul 29, 2006·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Andrew C NyborgTodd E Golde
Sep 18, 2004·The Biochemical Journal·Samuel G CrawshawStephen High
May 17, 2008·The Journal of Biological Chemistry·Alexandra ToliaBart De Strooper
May 23, 2007·The Journal of Biological Chemistry·Saravanakumar NarayananMichael S Wolfe
Oct 13, 2007·The Journal of Biological Chemistry·Lawrence G IbenJeremy H Toyn
Jan 1, 2021·Frontiers in Cardiovascular Medicine·Alkmini A Papadopoulou, Regina Fluhrer

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.