Characterization of angiotensin I-converting enzyme from anterior gills of the mangrove crab Ucides cordatus
| Autor: | Maria A. Juliano, Luiz Juliano, Paulo Bandiera Paiva, F.P. Zanotto, Regina F. Nogueira, Patrícia Alessandra Bersanetti, Adriana K. Carmona, Marcelo F. Marcondes |
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| Rok vydání: | 2015 |
| Předmět: |
Gene isoform
Gills animal structures Metallopeptidase Brachyura Characterization Bradykinin Angiotensin-Converting Enzyme Inhibitors Biology Peptidyl-Dipeptidase A Biochemistry FRET peptides Substrate Specificity chemistry.chemical_compound Affinity chromatography Lisinopril Structural Biology Enzyme Stability medicine Animals Molecular Biology Phylogeny chemistry.chemical_classification Crab gill Hydrolysis Temperature food and beverages Substrate (chemistry) General Medicine Hydrogen-Ion Concentration ACE isoform Enzyme Activation Enzyme chemistry ACE inhibitor medicine.drug |
| Zdroj: | International Journal of Biological Macromolecules. 74:304-309 |
| ISSN: | 0141-8130 |
| DOI: | 10.1016/j.ijbiomac.2014.12.036 |
| Popis: | Angiotensin I-converting enzyme (ACE) is a well-known metallopeptidase that is found in vertebrates, invertebrates and bacteria. We isolated from the anterior gill of the crab Ucides cordatus an isoform of ACE, here named crab-ACE, which presented catalytic properties closely resembling to those of mammalian ACE. The enzyme was purified on Sepharose-lisinopril affinity chromatography to apparent homogeneity and a band of about 72 kDa could be visualized after silver staining and Western blotting. Assays performed with fluorescence resonance energy transfer (FRET) selective ACE substrates Abz-FRK(Dnp)P-OH, Abz-SDK(Dnp)P-OH and Abz-LFK(Dnp)-OH, allowed us to verify that crab-ACE has hydrolytic profile very similar to that of the ACE C-domain. In addition, we observed that crab-ACE can hydrolyze the ACE substrates, angiotensin I and bradykinin. The enzyme was strongly inhibited by the specific ACE inhibitor lisinopril ( K i of 1.26 nM). However, in contrast to other ACE isoforms, crab-ACE presented a very particular optimum pH, being the substrate Abz-FRK(Dnp)-P-OH hydrolyzed efficiently at pH 9.5. Other interesting characteristic of crab-ACE was that the maximum hydrolytic activity was reached at around 45 °C. The description of an ACE isoform in Ucides cordatus is challenging and may contribute to a better understanding of the biochemical function of this enzyme in invertebrates. |
| Databáze: | OpenAIRE |
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