scholarly journals In vitro binding of Sorghum bicolor transcription factors ABI4 and ABI5 to a conserved region of a GA 2-OXIDASE promoter: possible role of this interaction in the expression of seed dormancy

2013 ◽  
Vol 64 (18) ◽  
pp. 5721-5735 ◽  
Author(s):  
Renata Cantoro ◽  
Carlos Daniel Crocco ◽  
Roberto Luis Benech-Arnold ◽  
María Verónica Rodríguez
Keyword(s):  
Transcription Factors ◽  
Sorghum Bicolor ◽  
Seed Dormancy ◽  
In Vitro Binding ◽  
Vitro Binding ◽  
Conserved Region

scholarly journals Phospholipase C–mediated hydrolysis of PIP2 releases ERM proteins from lymphocyte membrane

2009 ◽  
Vol 184 (3) ◽  
pp. 451-462 ◽  
Author(s):  
Jian-Jiang Hao ◽  
Yin Liu ◽  
Michael Kruhlak ◽  
Karen E. Debell ◽  
Barbara L. Rellahan ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Phospholipase C ◽  
Membrane Association ◽  
Erm Proteins ◽  
Cytoskeletal Remodeling ◽  
In Vitro Binding ◽  
Vitro Binding ◽  
Hydrolysis Of

Mechanisms controlling the disassembly of ezrin/radixin/moesin (ERM) proteins, which link the cytoskeleton to the plasma membrane, are incompletely understood. In lymphocytes, chemokine (e.g., SDF-1) stimulation inactivates ERM proteins, causing their release from the plasma membrane and dephosphorylation. SDF-1–mediated inactivation of ERM proteins is blocked by phospholipase C (PLC) inhibitors. Conversely, reduction of phosphatidylinositol 4,5-bisphosphate (PIP2) levels by activation of PLC, expression of active PLC mutants, or acute targeting of phosphoinositide 5-phosphatase to the plasma membrane promotes release and dephosphorylation of moesin and ezrin. Although expression of phosphomimetic moesin (T558D) or ezrin (T567D) mutants enhances membrane association, activation of PLC still relocalizes them to the cytosol. Similarly, in vitro binding of ERM proteins to the cytoplasmic tail of CD44 is also dependent on PIP2. These results demonstrate a new role of PLCs in rapid cytoskeletal remodeling and an additional key role of PIP2 in ERM protein biology, namely hydrolysis-mediated ERM inactivation.

scholarly journals Dynamics of Ankyrin-containing Complexes in Chicken Embryonic Erythroid Cells: Role of Phosphorylation

2001 ◽  
Vol 12 (12) ◽  
pp. 3864-3874 ◽  
Author(s):  
Sourav Ghosh ◽  
John V. Cox
Keyword(s):  
Complex Formation ◽  
Anion Exchanger ◽  
Erythroid Cells ◽  
Binding Studies ◽  
Phosphatase Inhibitors ◽  
In Vitro Binding ◽  
Vitro Binding

Chicken erythroid ankyrin undergoes a fairly rapid cycle of cytoskeletal association, dissociation, and turnover. In addition, the cytoskeletal association of ankyrin is regulated by phosphorylation. Treatment of erythroid cells with serine and threonine phosphatase inhibitors stimulated the hyperphosphorylation of the 225- and 205-kDa ankyrin isoforms, and dissociated the bulk of these isoforms from cytoskeletal spectrin. In vitro binding studies have shown that this dissociation of ankyrin from spectrin in vivo can be attributed to a reduced ability of hyperphosphorylated ankyrin to bind spectrin. Interestingly, a significant fraction of detergent insoluble ankyrin accumulates in a spectrin-independent pool. At least some of this spectrin-independent pool of ankyrin is complexed with the AE1 anion exchanger, and the solubility properties of this pool are also regulated by phosphorylation. Treatment of cells with serine and threonine phosphatase inhibitors had no effect on ankyrin/AE1 complex formation. However, these inhibitors were sufficient to shift ankyrin/AE1 complexes from the detergent insoluble to the soluble pool. These analyses, which are the first to document the in vivo consequences of ankyrin phosphorylation, indicate that erythroid ankyrin-containing complexes can undergo dynamic rearrangements in response to changes in phosphorylation.

Codon 12 Region of Mouse K-ras Gene Is the Site for in vitro Binding of Transcription Factors GATA-6 and NF-Y

2005 ◽  
Vol 70 (10) ◽  
pp. 1180-1184 ◽  
Author(s):  
E. V. Gorshkova ◽  
V. I. Kaledin ◽  
V. F. Kobzev ◽  
T. I. Merkulova
Keyword(s):  
Transcription Factors ◽  
Ras Gene ◽  
In Vitro Binding ◽  
Vitro Binding

scholarly journals The Development of Novel In Vitro Binding Assays to Further Elucidate the Role of tRNAs in Protein Synthesis

10.5772/21822 ◽  
2011 ◽  
Author(s):  
Anthony J. ◽  
Jr. ◽  
Suzanna Ellzey ◽  
Devin McDougald ◽  
Crystal Serrano
Keyword(s):  
Protein Synthesis ◽  
Binding Assays ◽  
In Vitro Binding ◽  
Vitro Binding

scholarly journals Molecular characterization of an alpha interferon receptor 1 subunit (IFNaR1) domain required for TYK2 binding and signal transduction.

1996 ◽  
Vol 16 (5) ◽  
pp. 2074-2082 ◽  
Author(s):  
H Yan ◽  
K Krishnan ◽  
J T Lim ◽  
L G Contillo ◽  
J J Krolewski
Keyword(s):  
Signal Transduction ◽  
Tyrosine Phosphorylation ◽  
Tyrosine Kinases ◽  
Binding Domain ◽  
Alpha Interferon ◽  
Binding Motif ◽  
In Vitro Binding ◽  
Vitro Binding

Binding of alpha interferon (IFNalpha) to its receptors induces rapid tyrosine phosphorylation of the receptor subunits IFNaR1 and IFNaR2, the TYK2 and JAK1 tyrosine kinases, and the Stat1 and Stat2 transcription factors. Previous studies have demonstrated that TYK2 directly and specifically binds to and tyrosine phosphorylates IFNaR1 in vitro. We now report a detailed analysis of the TYK2 binding domain on the IFNaR1 subunit. First, we used an in vitro binding assay to identify the TYK2 binding motif in IFNaR1 as well as the critical residues within this region. The most striking feature is the importance of a number of hydrophobic and acidic residues. A minor role is also ascribed to a region resembling the proline-rich "box 1" sequence. In addition, mutations which disrupt in vitro binding also disrupt the coimmunoprecipitation of the receptor and TYK2. We also provide direct evidence that the binding region is both necessary and sufficient to activate TYK2 in vivo. Specifically, mutations in the binding domain act in a dominant-negative fashion to inhibit the IFNalpha-induced tyrosine phosphorylation of TYK2 and Stat2. Further, introduction of dimerized glutathione S-transferase-IFNaR1 fusion proteins into permeabilized cells is sufficient to induce phosphorylation of TYK2 and the receptor, confirming the role of the binding domain in IFNalpha signal transduction. These studies provide clues to the sequences determining the specificity of the association between JAK family tyrosine kinases and cytokine receptors as well as the functional role of these kinases in cytokine signal transduction.

Discovery of New AKT1 Inhibitors by Combination of In silico Structure Based Virtual Screening Approaches and Biological Evaluations

2019 ◽  
Author(s):  
Filip Fratev ◽  
Denisse A. Gutierrez ◽  
Renato J. Aguilera ◽  
suman sirimulla
Keyword(s):  
Virtual Screening ◽  
Success Rate ◽  
Protein Interactions ◽  
In Silico ◽  
Biological Data ◽  
In Vitro Binding ◽  
Vitro Binding ◽  
Screening Protocol ◽  
Screening Approaches

AKT1 is emerging as a useful target for treating cancer. Herein, we discovered a new set of ligands that inhibit the AKT1, as shown by in vitro binding and cell line studies, using a newly designed virtual screening protocol that combines structure-based pharmacophore and docking screens. Taking together with the biological data, the combination of structure based pharamcophore and docking methods demonstrated reasonable success rate in identifying new inhibitors (60-70%) proving the success of aforementioned approach. A detail analysis of the ligand-protein interactions was performed explaining observed activities.<br>

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