Photocontrol of Tyrosine Phosphorylation in Mammalian Cells via Genetic Encoding of Photocaged Tyrosine

2012 ◽  
Vol 134 (29) ◽  
pp. 11912-11915 ◽  
Author(s):  
Eyal Arbely ◽  
Jessica Torres-Kolbus ◽  
Alexander Deiters ◽  
Jason W. Chin
Keyword(s):  
Tyrosine Phosphorylation ◽  
Mammalian Cells ◽  
Genetic Encoding

Tyrosine phosphorylation controls nuclear localization and transcriptional activity of Ssdp1 in mammalian cells

2008 ◽  
Vol 103 (6) ◽  
pp. 1856-1865 ◽  
Author(s):  
Ipsita Dey-Guha ◽  
Nasir Malik ◽  
Renaud Lesourne ◽  
Paul E. Love ◽  
Heiner Westphal
Keyword(s):  
Tyrosine Phosphorylation ◽  
Nuclear Localization ◽  
Mammalian Cells ◽  
Transcriptional Activity

Photoactivation of Innate Immunity Receptor TLR8 in Live Mammalian Cells by Genetic Encoding of Photocaged Tyrosine

ChemBioChem ◽  
2021 ◽  
Author(s):  
Yi Yang ◽  
Shuchen Luo ◽  
Jian Huang ◽  
Yu Xiao ◽  
Yixuan Fu ◽  
...  
Keyword(s):  
Innate Immunity ◽  
Mammalian Cells ◽  
Genetic Encoding

scholarly journals Functional genetic encoding of sulfotyrosine in mammalian cells

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Xinyuan He ◽  
Yan Chen ◽  
Daisy Guiza Beltran ◽  
Maia Kelly ◽  
Bin Ma ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Biochemical Characterization ◽  
Trna Synthetase ◽  
Functional Verification ◽  
Cellular Processes ◽  
Genetic Encoding ◽  
Efficient Expression

Abstract Protein tyrosine O-sulfation (PTS) plays a crucial role in extracellular biomolecular interactions that dictate various cellular processes. It also involves in the development of many human diseases. Regardless of recent progress, our current understanding of PTS is still in its infancy. To promote and facilitate relevant studies, a generally applicable method is needed to enable efficient expression of sulfoproteins with defined sulfation sites in live mammalian cells. Here we report the engineering, in vitro biochemical characterization, structural study, and in vivo functional verification of a tyrosyl-tRNA synthetase mutant for the genetic encoding of sulfotyrosine in mammalian cells. We further apply this chemical biology tool to cell-based studies on the role of a sulfation site in the activation of chemokine receptor CXCR4 by its ligand. Our work will not only facilitate cellular studies of PTS, but also paves the way for economical production of sulfated proteins as therapeutic agents in mammalian systems.

scholarly journals The Drosophila insulin receptor activates multiple signaling pathways but requires insulin receptor substrate proteins for DNA synthesis.

1996 ◽  
Vol 16 (5) ◽  
pp. 2509-2517 ◽  
Author(s):  
L Yenush ◽  
R Fernandez ◽  
M G Myers ◽  
T C Grammer ◽  
X J Sun ◽  
...  
Keyword(s):  
Insulin Receptor ◽  
Tyrosine Phosphorylation ◽  
Human Insulin ◽  
Mammalian Cells ◽  
Insulin Response ◽  
Mitogen Activated Protein Kinase ◽  
Insulin Receptor Substrate ◽  
Human Insulin Receptor ◽  
Irs Proteins ◽  
Drosophila Insulin Receptor

The Drosophila insulin receptor (DIR) contains a 368-amino-acid COOH-terminal extension that contains several tyrosine phosphorylation sites in YXXM motifs. This extension is absent from the human insulin receptor but resembles a region in insulin receptor substrate (IRS) proteins which binds to the phosphatidylinositol (PI) 3-kinase and mediates mitogenesis. The function of a chimeric DIR containing the human insulin receptor binding domain (hDIR) was investigated in 32D cells, which contain few insulin receptors and no IRS proteins. Insulin stimulated tyrosine autophosphorylation of the human insulin receptor and hDIR, and both receptors mediated tyrosine phosphorylation of Shc and activated mitogen-activated protein kinase. IRS-1 was required by the human insulin receptor to activate PI 3-kinase and p70s6k, whereas hDIR associated with PI 3-kinase and activated p70s6k without IRS-1. However, both receptors required IRS-1 to mediate insulin-stimulated mitogenesis. These data demonstrate that the DIR possesses additional signaling capabilities compared with its mammalian counterpart but still requires IRS-1 for the complete insulin response in mammalian cells.

scholarly journals Tyrosine phosphorylation and activation of homologous protein kinases during oocyte maturation and mitogenic activation of fibroblasts.

1991 ◽  
Vol 11 (5) ◽  
pp. 2517-2528 ◽  
Author(s):  
J Posada ◽  
J Sanghera ◽  
S Pelech ◽  
R Aebersold ◽  
J A Cooper
Keyword(s):  
Cell Cycle ◽  
Map Kinase ◽  
Tyrosine Phosphorylation ◽  
Oocyte Maturation ◽  
Mammalian Cells ◽  
Map Kinases ◽  
Kinase Activity ◽  
Mitotic Cell ◽  
Meiotic Maturation ◽  
Sea Star

Meiotic maturation of Xenopus and sea star oocytes involves the activation of a number of protein-serine/threonine kinase activities, including a myelin basic protein (MBP) kinase. A 44-kDa MBP kinase (p44mpk) purified from mature sea star oocytes is shown here to be phosphorylated at tyrosine. Antiserum to purified sea star p44mpk was used to identify antigenically related proteins in Xenopus oocytes. Two tyrosine-phosphorylated 42-kDa proteins (p42) were detected with this antiserum in Xenopus eggs. Xenopus p42 chromatographs with MBP kinase activity on a Mono Q ion-exchange column. Tyrosine phosphorylation of Xenopus p42 approximately parallels MBP kinase activity during meiotic maturation. These results suggest that related MBP kinases are activated during meiotic maturation of Xenopus and sea star oocytes. Previous studies have suggested that Xenopus p42 is related to the mitogen-activated protein (MAP) kinases of culture mammalian cells. We have cloned a MAP kinase relative from a Xenopus ovary cDNA library and demonstrate that this clone encodes the Xenopus p42 that is tyrosine phosphorylated during oocyte maturation. Comparison of the sequences of Xenopus p42 and a rat MAP kinase (ERK1) and peptide sequences from sea star p44mpk indicates that these proteins are close relatives. The family members appear to be tyrosine phosphorylated, and activated, in different contexts, with the murine MAP kinase active during the transition from quiescence to the G1 stage of the mitotic cell cycle and the sea star and Xenopus kinases being active during M phase of the meiotic cell cycle.

Tyrosine phosphorylation of the Rab24 GTPase in cultured mammalian cells

2003 ◽  
Vol 312 (3) ◽  
pp. 670-675 ◽  
Author(s):  
Jane Ding ◽  
Gwendolyn Soule ◽  
Jean H Overmeyer ◽  
William A Maltese
Keyword(s):  
Tyrosine Phosphorylation ◽  
Mammalian Cells

Genetic Encoding of Caged Cysteine and Caged Homocysteine in Bacterial and Mammalian Cells

ChemBioChem ◽  
2014 ◽  
Vol 15 (12) ◽  
pp. 1793-1799 ◽  
Author(s):  
Rajendra Uprety ◽  
Ji Luo ◽  
Jihe Liu ◽  
Yuta Naro ◽  
Subhas Samanta ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Genetic Encoding

scholarly journals Further evidence that the tyrosine phosphorylation of glycogen synthase kinase-3 (GSK3) in mammalian cells is an autophosphorylation event

2004 ◽  
Vol 377 (1) ◽  
pp. 249-255 ◽  
Author(s):  
Adam COLE ◽  
Sheelagh FRAME ◽  
Philip COHEN
Keyword(s):  
Tyrosine Phosphorylation ◽  
Mammalian Cells ◽  
Protein Tyrosine Kinase ◽  
Kinase Inhibitor ◽  
Glycogen Synthase ◽  
Glycogen Synthase Kinase ◽  
Hek 293 ◽  
293 Cells ◽  
General Protein ◽  
The Stability

Phosphorylation of the endogenous GSK3α (glycogen synthase kinase-3α) at Tyr279 and GSK3β at Tyr216 was suppressed in HEK-293 or SH-SY5Y cells by incubation with pharmacological inhibitors of GSK3, but not by an Src-family inhibitor, 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP2), or a general protein tyrosine kinase inhibitor (genistein). GSK3β transfected into HEK-293 cells or Escherichia coli became phosphorylated at Tyr216, but catalytically inactive mutants did not. GSK3β expressed in insect Sf 21 cells or E. coli was extensively phosphorylated at Tyr216, but the few molecules lacking phosphate at this position could autophosphorylate at Tyr216in vitro after incubation with MgATP. The rate of autophosphorylation was unaffected by dilution and was suppressed by the GSK3 inhibitor kenpaullone. Wild-type GSK3β was unable to catalyse the tyrosine phosphorylation of catalytically inactive GSK3β lacking phosphate at Tyr216. Our results indicate that the tyrosine phosphorylation of GSK3 is an intramolecular autophosphorylation event in the cells that we have studied and that this modification enhances the stability of the enzyme.

scholarly journals Genetic Encoding of A Nonhydrolyzable Phosphotyrosine Analog in Mammalian Cells

2021 ◽  
Author(s):  
Wei Niu ◽  
Jiantao Guo ◽  
Xinyuan He ◽  
Bin Ma ◽  
Yan Chen
Keyword(s):  
Tyrosine Phosphorylation ◽  
Mammalian Cells ◽  
Critical Role ◽  
Protein Tyrosine Phosphorylation ◽  
Protein Tyrosine ◽  
Practical Applications ◽  
Cellular Processes ◽  
Cellular Events ◽  
Biological Studies ◽  
Model Protein

Protein tyrosine phosphorylation plays a critical role in signal transduction and the regulation of many cellular processes. It is of great significance to understand the underlying regulatory mechanism of particular tyrosine phosphorylation events. Here we report the genetic incorporation of a phosphotyrosine (pTyr) analog, p-carboxymethyl-L-phenylalanine (CMF), into proteins in mammalian cells. This nonhydrolyzable pTyr analog can facilitate biological studies by removing complications caused by the dynamic interconversion between the phosphorylated and non-phosphorylated isoforms of a protein. The developed methodology was demonstrated by using the human signal transducer and activator of transcription-1 (STAT1) as a model protein for homogeneous and defined incorporation of CMF. This tool will greatly enhance our capability to study protein tyrosine phosphorylation-associated biomolecular and cellular events, and enhance biomedical research that target protein tyrosine phosphorylation, which will have a broad impact to both fundamental studies and practical applications.

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