Preparation and Biological Activity of Molecular Probes to Identify and Analyze Jasmonic Acid-binding Proteins

2004 ◽  
Vol 68 (7) ◽  
pp. 1461-1466 ◽  
Author(s):  
Yusuke JIKUMARU ◽  
Tadao ASAMI ◽  
Hideharu SETO ◽  
Shigeo YOSHIDA ◽  
Tadashi YOKOYAMA ◽  
...  
Keyword(s):  
Biological Activity ◽  
Jasmonic Acid ◽  
Binding Proteins ◽  
Molecular Probes

scholarly journals Three distinct anti-allergic drugs, amlexanox, cromolyn and tranilast, bind to S100A12 and S100A13 of the S100 protein family

1999 ◽  
Vol 338 (3) ◽  
pp. 583-589 ◽  
Author(s):  
Tsuyoshi SHISHIBORI ◽  
Yuhta OYAMA ◽  
Osamu MATSUSHITA ◽  
Kayoko YAMASHITA ◽  
Hiromi FURUICHI ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Calcium Binding ◽  
Binding Proteins ◽  
S100 Protein ◽  
Reversed Phase ◽  
Molecular Probes ◽  
Calcium Binding Proteins ◽  
Ige Mediated ◽  
Human And Mouse

To investigate the roles of calcium-binding proteins in degranulation, we used three anti-allergic drugs, amlexanox, cromolyn and tranilast, which inhibit IgE-mediated degranulation of mast cells, as molecular probes in affinity chromatography. All of these drugs, which have different structures but similar function, scarcely bound to calmodulin in bovine lung extract, but bound to the same kinds of calcium-binding proteins, such as the 10-kDa proteins isolated in this study, calcyphosine and annexins I–V. The 10-kDa proteins obtained on three drug-coupled resins and on phenyl-Sepharose were analysed by reversed-phase HPLC. It was found that two characteristic 10-kDa proteins, one polar and one less polar, were bound with all three drugs, although S100A2 (S100L), of the S100 family, was bound with phenyl-Sepharose. The cDNA and deduced amino acid sequence proved our major polar protein to be identical with the calcium-binding protein in bovine amniotic fluid (CAAF1, S100A12). The cDNA and deduced amino acid sequence of the less-polar protein shared 95% homology with human and mouse S100A13. In addition, it was demonstrated that the native S100A12 and recombinant S100A12 and S100A13 bind to immobilized amlexanox. On the basis of these findings, we speculate that the three anti-allergic drugs might inhibit degranulation by binding with S100A12 and S100A13.

scholarly journals N1-Propargylguanosine Modified mRNA Cap Analogs: Synthesis, Reactivity, and Applications to the Study of Cap-Binding Proteins

Molecules ◽  
2019 ◽  
Vol 24 (10) ◽  
pp. 1899
Author(s):  
Michal Kopcial ◽  
Blazej A. Wojtczak ◽  
Renata Kasprzyk ◽  
Joanna Kowalska ◽  
Jacek Jemielity
Keyword(s):  
Protein Interactions ◽  
Binding Proteins ◽  
Molecular Probes ◽  
Translation Initiation Factor ◽  
Initiation Factor ◽  
Binding Assays ◽  
Fluorescent Properties ◽  
Eukaryotic Translation ◽  
Eukaryotic Translation Initiation ◽  
Fluorescent Tags

The mRNA 5′ cap consists of N7-methylguanosine bound by a 5′,5′-triphosphate bridge to the first nucleotide of the transcript. The cap interacts with various specific proteins and participates in all key mRNA-related processes, which may be of therapeutic relevance. There is a growing demand for new biophysical and biochemical methods to study cap–protein interactions and identify the factors which inhibit them. The development of such methods can be aided by the use of properly designed fluorescent molecular probes. Herein, we synthesized a new class of m7Gp3G cap derivatives modified with an alkyne handle at the N1-position of guanosine and, using alkyne-azide cycloaddition, we functionalized them with fluorescent tags to obtain potential probes. The cap derivatives and probes were evaluated in the context of two cap-binding proteins, eukaryotic translation initiation factor (eIF4E) and decapping scavenger (DcpS). Biochemical and biophysical studies revealed that N1-propargyl moiety did not significantly disturb cap–protein interaction. The fluorescent properties of the probes turned out to be in line with microscale thermophoresis (MST)-based binding assays.

scholarly journals Modulation of the Biological Activity of a Tobacco LTP1 by Lipid Complexation

2004 ◽  
Vol 15 (11) ◽  
pp. 5047-5052 ◽  
Author(s):  
Nathalie Buhot ◽  
Eric Gomès ◽  
Marie-Louise Milat ◽  
Michel Ponchet ◽  
Didier Marion ◽  
...  
Keyword(s):  
Biological Activity ◽  
Plasma Membrane ◽  
Jasmonic Acid ◽  
Plant Disease Resistance ◽  
Plasma Membranes ◽  
Biological Properties ◽  
Membrane Receptors ◽  
Lipid Transfer ◽  
Acid Complex ◽  
Ligand Interaction

Plant lipid transfer proteins (LTPs) are small, cysteine-rich proteins secreted into the extracellular space. They belong to the pathogenesis-related proteins (PR-14) family and are believed to be involved in several physiological processes including plant disease resistance, although their precise biological function is still unknown. Here, we show that a recombinant tobacco LTP1 is able to load fatty acids and jasmonic acid. This LTP1 binds to specific plasma membrane sites, previously characterized as elicitin receptors, and is shown to be involved in the activation of plant defense. The biological properties of this LTP1 were compared with those of LTP1-linolenic and LTP1-jasmonic acid complexes. The binding curve of the LTP1-linolenic acid complex to purified tobacco plasma membranes is comparable to the curve obtained with LTP1. In contrast, the LTP1-jasmonic acid complex shows a strongly increased interaction with the plasma membrane receptors. Treatment of tobacco plants with LTP1-jasmonic acid resulted in an enhancement of resistance toward Phytophthora parasitica. These effects were absent upon treatment with LTP1 or jasmonic acid alone. This work presents the first evidence for a biological activity of a LTP1 and points out the crucial role of protein-specific lipophilic ligand interaction in the modulation of the protein activity.

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