N-domain angiotensin-I converting enzyme is expressed in immortalized mesangial, proximal tubule and collecting duct cells
| Autor: | Giovana Seno Di Marco, Renato A. Mortara, Beata Marie Redublo Quinto, Maria Claudina Camargo de Andrade, Pamella Huey Mei Wang, Carlos P. Vio, Dulce Elena Casarini |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Lysis
Somatic cell Size-exclusion chromatography Nephron Peptidyl-Dipeptidase A Biochemistry Gene Expression Regulation Enzymologic Madin Darby Canine Kidney Cells Substrate Specificity Mice Dogs Structural Biology medicine Animals Humans Molecular Biology chemistry.chemical_classification urogenital system Captopril General Medicine Chromatography Ion Exchange Protein Structure Tertiary Rats medicine.anatomical_structure Enzyme chemistry Mesangial Cells Rabbits Oligopeptides Duct (anatomy) Intracellular medicine.drug |
| Zdroj: | International Journal of Biological Macromolecules Artículos CONICYT CONICYT Chile instacron:CONICYT |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|