Purification of a Src family tyrosine protein kinase from bovine retinas
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Deisy Perdomo
Abstract
Since tyrosine phosphorylation appears to play important functions in photoreceptor cells, we searched here for retinal nonreceptor tyrosine kinases of the Src family. We demonstrated that Src family tyrosine kinases were present in the cytosolic fraction of extracted bovine retinas. A Src family tyrosine kinase with an apparent molecular mass of about 62 kDa was purified to homogeneity from the soluble fraction of dark-adapted bovine retinas after three consecutive purification steps: ω-aminooctyl-agarose hydrophobic chromatography, Cibacron blue 3GA-agarose pseudo-affinity chromatography, and α-casein-agarose affinity chromatography. The purified protein was subjected to N-terminal amino acid sequencing and the sequence Gly-Ile-Ile-Lys-Ser-Glu-Glu was obtained, which displayed homology with the first seven residues of the Src family tyrosine kinase c-Yes from Bos taurus (Gly-Cys-Ile-Lys-Ser-Lys-Glu). Although the cytosolic fraction from dark-adapted retinas contained tyrosine kinases of the Src family capable of phosphorylating the α-subunit of transducin, which is the heterotrimeric G protein involved in phototransduction, the purified tyrosine kinase was not capable of using transducin as a substrate. The cellular role of this retinal Src family member remains to be found.
Funding source: Fondo Nacional de Ciencia Tecnología e Innovación
Award Identifier / Grant number: S1-2000000514
Acknowledgments
This work was supported in part by a grant from FONACIT, Caracas, Venezuela [grant number S1-2000000514]. We would like to thank Dr. Johan Hoebeke (Université Louis Pasteur Strasbourg, Strasbourg, France) for generously performing the amino acid sequencing.
Author contribution: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: This study was funded by FONACIT, Caracas, Venezuela [grant number S1-2000000514].
Conflict of interest statement: The authors declare no conflicts of interest regarding this article.
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© 2020 Walter de Gruyter GmbH, Berlin/Boston
Artikel in diesem Heft
- Frontmatter
- Research Articles
- Ultra-high-performance liquid chromatography – high-resolution mass spectrometry profiling and hepatoprotective activity of purified saponin and flavonoid fractions from the aerial parts of wild spinach (Chenopodium bonus-henricus L.)
- Purification of a Src family tyrosine protein kinase from bovine retinas
- Phytochemical, antibacterial, antioxidant and cytoxicity investigation of Tarenna grandiflora
- Cytogenetic and oxidative effects of three lichen extracts on human peripheral lymphocytes
- Cardenolides and pentacyclic triterpenes isolated from Acokanthera oblongifolia leaves: their biological activities with molecular docking study
- Assessment of Cota altissima (L.) J. Gay for phytochemical composition and antioxidant, anti-inflammatory, antidiabetic and antimicrobial activities
- Anticancer effects of novel NSAIDs derivatives on cultured human glioblastoma cells
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Artikel in diesem Heft
- Frontmatter
- Research Articles
- Ultra-high-performance liquid chromatography – high-resolution mass spectrometry profiling and hepatoprotective activity of purified saponin and flavonoid fractions from the aerial parts of wild spinach (Chenopodium bonus-henricus L.)
- Purification of a Src family tyrosine protein kinase from bovine retinas
- Phytochemical, antibacterial, antioxidant and cytoxicity investigation of Tarenna grandiflora
- Cytogenetic and oxidative effects of three lichen extracts on human peripheral lymphocytes
- Cardenolides and pentacyclic triterpenes isolated from Acokanthera oblongifolia leaves: their biological activities with molecular docking study
- Assessment of Cota altissima (L.) J. Gay for phytochemical composition and antioxidant, anti-inflammatory, antidiabetic and antimicrobial activities
- Anticancer effects of novel NSAIDs derivatives on cultured human glioblastoma cells
- In vitro chemotherapeutic and antiangiogenic properties of cardenolides from Acokanthera oblongifolia (Hochst.) Codd
