Biosynthesis, processing, trafficking, and enzymatic activity of mouse neprilysin 2.

Molecular and Cellular Biochemistry
Kentaro Oh-hashiKazutoshi Kiuchi

Abstract

Neprilysin 2 (NEP2) has been recently identified as a new member of the M13 subfamily of zinc-dependent metalloproteases and shares a highly homologous amino acid sequence with neprilysin (EC 3.4.24.11, NEP). NEP2 has been reported to exist as membrane-bound and soluble secreted variants. To investigate mechanisms of regulating NEP2 activity, we developed a simple and sensitive method for measuring NEP2 activity using synthetic substrates with a fluorescent probe. NEP2 only cleaved Suc-Ala-Ala-Phe-AMC, while NEP cleaved both Dansyl-D-Ala-Gly-p-nitro-Phe-Gly and Suc-Ala-Ala-Phe-AMC. Using HEK293 cells stably expressing mouse NEP2, we evaluated the effects of various reagents affecting post-translational modification and protein trafficking on extracellular NEP2 activity secreted into the culture medium. Inhibition of N-glycosylation by tunicamycin reduced both the enzymatic activity of extracellular NEP2 and the molecular size of intracellular NEP2. Disruption of the Golgi apparatus with brefeldin A markedly reduced extracellular NEP2 activity in parallel with intracellular NEP2 protein level in HEK293 cells. In contrast, the cytoskeleton disrupting reagents, nocodazole and cytochalasin B barely affected NEP2 activity. Two disti...Continue Reading

References

Jul 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·S LeeC M Redman
Apr 14, 1987·Biochemical and Biophysical Research Communications·B MalfroyW J Henzel
Apr 12, 2000·Proceedings of the National Academy of Sciences of the United States of America·S Kiryu-SeoH Kiyama
May 18, 2000·Biochemical and Biophysical Research Communications·T OuimetO Tanja
Feb 27, 2001·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·A J TurnerD Coates
Mar 10, 2001·Trends in Pharmacological Sciences·J Bralet, J C Schwartz
Apr 20, 2002·The Biochemical Journal·Christiane RoseTanja Ouimet
Dec 25, 2002·European Journal of Pharmacology·Takeyuki YatsuSusumu Satoh
Dec 20, 2003·Cell Death and Differentiation·S Oyadomari, M Mori
Jun 23, 2004·The Journal of Biological Chemistry·Ashish K Sharma, Baerbel Rohrer
Mar 19, 2005·Journal of Neurochemistry·Flávia R CarreñoEmer S Ferro
Jul 12, 2005·Biochemical and Biophysical Research Communications·Kentaro Oh-hashiKazutoshi Kiuchi

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Citations

Nov 30, 2011·Journal of Neurochemistry·Natalia N NalivaevaAnthony J Turner
Jan 13, 2011·The American Journal of Pathology·Daniel HafezRobert A Marr
Dec 1, 2010·Respiratory Research·Soshi HashimotoSatoru Hashimoto

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