| Full text | |
| Author(s): |
Mei Wang, Pamella Huey
[1]
;
Andrade, Maria Claudina
[1]
;
Redublo Quinto, Beata Marie
[1]
;
Di Marco, Giovana
[1]
;
Mortara, Renato Arruda
[2]
;
Vio, Carlos P.
[3]
;
Casarini, Dulce Elena
[1]
Total Authors: 7
|
| Affiliation: | [1] Univ Fed Sao Paulo UNIFESP, Disciplina Nefrol, Dept Med, BR-04023900 Sao Paulo - Brazil
[2] Univ Fed Sao Paulo UNIFESP, Disciplina Parasitol, Dept Microimunoparasitol, BR-04023900 Sao Paulo - Brazil
[3] PUC, Dept Fisiol, Santiago - Chile
Total Affiliations: 3
|
| Document type: | Review article |
| Source: | International Journal of Biological Macromolecules; v. 72, p. 380-390, JAN 2015. |
| Web of Science Citations: | 1 |
| 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. (AU) | |
| FAPESP's process: | 02/13290-2 - Angiotensin I: converting enzyme isoform (90 kDa), potential genetic marker of hypertension: processing, molecular and functional characterization, and genetic segregation |
| Grantee: | Dulce Elena Casarini |
| Support Opportunities: | Research Projects - Thematic Grants |