N-domain angiotensin-I converting enzyme is expressed in immortalized mesangial, proximal tubule and collecting duct cells

N-domain angiotensin-I converting enzyme is expressed in immortalized mesangial, proximal tubule and collecting duct cells

Author Mei Wang, Pamella Huey Autor UNIFESP Google Scholar
Andrade, Maria Claudina Autor UNIFESP Google Scholar
Redublo Quinto, Beata Marie Autor UNIFESP Google Scholar
Di Marco, Giovana Autor UNIFESP Google Scholar
Mortara, Renato Arruda Autor UNIFESP Google Scholar
Vio, Carlos P. Google Scholar
Casarini, Dulce Elena Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
PUC
Abstract Somatic ACE (sACE) is found in glomerulus, proximal tubule and excreted in urine. We hypothesized that N-domain ACE can also be found at these sites. ACE profile was analyzed in mesangial (IMC), proximal (LLC-PK1), distal tubule (MDCK) and collecting duct (IMCD) cells. Cell lysate and culture medium were submitted to gel filtration chromatography, which separated two peaks with ACE activity from cells and medium, except from distal tubule. the first had a high molecular weight and the second, a lower one (65 kDa; N-domain ACE). We focused on N-domain ACE purification and characterization from LLC-PK1. Total LLC-PK1 N-domain ACE purification was achieved by ion-exchange chromatography, which presented only one peak with ACE activity, denominated ACE(int2A).ACE(int2A) activity was influenced by pH, NaCl and temperature. the purified enzyme was inhibited by Captopril and hydrolyzed AngI, Ang1-7 and AcSDKP. Its ability to hydrolyze AcSDKP characterized it as an N-domain ACE. ACE(int2A) also presented high amino acid sequence homology with the N-terminal part of sACE from mouse, rat, human and rabbit. the presence of secreted and intracellular N-domain ACE and sACE in IMC, LLC-PK1 and IMCD cells confirmed our studies along the nephron. We identified, purified and characterized N-domain ACE from LLC-PK1. (C) 2014 Elsevier B.V. All rights reserved.
Keywords Angiotensin-I converting enzyme
N-domain ACE
Proximal tubule cells
LLC-PK1
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
FONDECYT
Grant number FAPESP: 02/13290-2
FONDECYT: 1050997
Date 2015-01-01
Published in International Journal of Biological Macromolecules. Amsterdam: Elsevier B.V., v. 72, p. 380-390, 2015.
ISSN 0141-8130 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 380-390
Origin http://dx.doi.org/10.1016/j.ijbiomac.2014.07.043
Access rights Closed access
Type Review
Web of Science ID WOS:000347577900051
URI http://repositorio.unifesp.br/handle/11600/38584

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