Identification of a crab gill FXYD2 protein and regulation of crab microsomal Na, K-ATPase activity by mammalian FXYD2 peptide
| Autor: | D. C. Masui, Jonas Perales, Rosa dos Prazeres Melo Furriel, Carlos Frederico Leite Fontes, Helena M. Scofano, Francisco A. Leone, Hector Barrabin, John Campbell McNamara, André Teixeira-Ferreira, E. C. C. Silva |
|---|---|
| Rok vydání: | 2012 |
| Předmět: |
Gill
Gills animal structures Callinectes Swine Biophysics Peptide Stimulation Biology Biochemistry Crustacea Microsomes Animals Humans Na+/K+-ATPase Phosphorylation Endogenous kinase chemistry.chemical_classification FXYD2 peptide Cell Biology biology.organism_classification Cyclic AMP-Dependent Protein Kinases Enzyme assay ENZIMAS Enzyme Gill microsome Euryhaline crab chemistry Na K-ATPase Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization biology.protein Electrophoresis Polyacrylamide Gel Sodium-Potassium-Exchanging ATPase |
| Zdroj: | Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual) Universidade de São Paulo (USP) instacron:USP |
| Popis: | This investigation discloses the recognition of an FXYD2 protein in a microsomal Na,K-ATPase preparation from the posterior gills of the blue crab, Callinectes danae, by a mammalian (rabbit) FXYD2 peptide specific antibody (γC(33)) and MALDI-TOF-TOF mass spectrometry techniques. This is the first demonstration of an invertebrate FXYD2 protein. The addition of exogenous pig FXYD2 peptide to the crab gill microsomal fraction stimulated Na,K-ATPase activity in a dose-dependent manner. Exogenous pig FXYD2 also considerably increased enzyme affinity for K(+), ATP and NH(4)(+). K(0.5) for Na(+) was unaffected. Exogenous pig FXYD2 increased the V(max) for stimulation of gill Na,K-ATPase activity by Na(+), K(+) and ATP, by 30% to 40%. The crab gill FXYD2 is phosphorylated by PKA, suggesting a regulatory function similar to that known for the mammalian enzyme. The PKA-phosphorylated pig FXYD2 peptide stimulated the crab gill Na,K-ATPase activity by 80%, about 2-fold greater than did the non-phosphorylated peptide. Stimulation by the PKC-phosphorylated pig FXYD2 peptide was minimal. These findings confirm the presence of an FXYD2 peptide in the crab gill Na,K-ATPase and demonstrate that this peptide plays an important role in regulating enzyme activity. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|