scholarly journals Investigation of 1-Methylcytosine as a Ligand in Gold(III) Complexes: Synthesis and Protein Interactions

Inorganics ◽  
2018 ◽  
Vol 7 (1) ◽  
pp. 1 ◽  
Author(s):  
James Beaton ◽  
Nicholas Farrell
Keyword(s):  
Fluorescence Quenching ◽  
Zinc Finger ◽  
Protein Interactions ◽  
Nucleocapsid Protein ◽  
1H Nmr Spectroscopy ◽  
Ionization Mass ◽  
Target Interaction ◽  
Self Association ◽  
A Charge ◽  
First Time

The HIV nucleocapsid protein NCp7 was previously shown to play a number of roles in the viral life cycle and was previously identified as a potential target for small molecule intervention. In this work, the synthesis of the previously unreported complexes [Au(dien)(1MeCyt)]3+, [Au(N-Medien)(1MeCyt)]3+, and [Au(dien)(Cyt)]3+ is detailed, and the interactions of these complexes with the models for NCp7 are described. The affinity for these complexes with the target interaction site, the “essential” tryptophan of the C-terminal zinc finger motif of NCp7, was investigated through the use of a fluorescence quenching assay and by 1H-NMR spectroscopy. The association of [Au(dien)(1MeCyt)]3+ as determined through fluorescence quenching is intermediate between the previously reported DMAP and 9-EtGua analogs, while the associations of [Au(N-Medien)(1MeCyt)]3+ and [Au(dien)(Cyt)]3+ are lower than the previously reported complexes. Additionally, NMR investigation shows that the self-association of relevant compounds is negligible. The specifics of the interaction with the C-terminal zinc finger were investigated by circular dichroism spectroscopy and electrospray-ionization mass spectrometry. The interaction is complete nearly immediately upon mixing, and the formation of AuxFn+ (x = 1, 2, or 4; F = apopeptide) concomitant with the loss of all ligands is observed. Additionally, oxidized dimerized peptide was observed for the first time as a product, indicating a reaction via a charge transfer mechanism.

scholarly journals Self-association of the erythroid transcription factor GATA-1 mediated by its zinc finger domains.

1995 ◽  
Vol 15 (5) ◽  
pp. 2448-2456 ◽  
Author(s):  
M Crossley ◽  
M Merika ◽  
S H Orkin
Keyword(s):  
Transcription Factors ◽  
Dna Binding ◽  
Zinc Finger ◽  
Protein Interactions ◽  
Physical Interaction ◽  
Erythroid Cells ◽  
Protein Protein Interactions ◽  
Self Association ◽  
Zinc Finger Region ◽  
In Erythroid Cells

GATA-1, the founding member of a distinctive family of transcription factors, is expressed predominantly in erythroid cells and participates in the expression of numerous erythroid cell-expressed genes. GATA-binding sites are found in the promoters and enhancers of globin and nonglobin erythroid genes as well as in the alpha- and beta-globin locus control regions. To elucidate how GATA-1 may function in a variety of regulatory contexts, we have examined its protein-protein interactions. Here we show that GATA-1 self-associates in solution and in whole-cell extracts and that the zinc finger region of the molecule is sufficient to mediate this interaction. This physical interaction can influence transcription, as GATA-1 self-association is able to recruit a transcriptionally active but DNA-binding-defective derivative of GATA-1 to promoter-bound GATA-1 and result in superactivation. Through in vitro studies with bacterially expressed glutathione S-transferase fusion proteins, we have localized the minimal domain required for GATA-1 self-association to 40 amino acid residues within the C-terminal zinc finger region. Finally, we have detected physical interaction of GATA-1 with other GATA family members (GATA-2 and GATA-3) also mediated through the zinc finger domain. These findings have broad implications for the involvement of GATA factors in transcriptional control. In particular, the interaction of GATA-1 with itself and with other transcription factors may facilitate its function at diverse promoters in erythroid cells and also serve to bring together, or stabilize, loops between distant regulatory elements, such as the globin locus control regions and downstream globin promoters. We suggest that the zinc finger region of GATA-1, and related proteins, is multifunctional and mediates not only DNA binding but also important protein-protein interactions.

scholarly journals Four Major Envelope Proteins of White Spot Syndrome Virus Bind To Form a Complex

2009 ◽  
Vol 83 (9) ◽  
pp. 4709-4712 ◽  
Author(s):  
Qing Zhou ◽  
Limei Xu ◽  
Hui Li ◽  
Yi-Peng Qi ◽  
Feng Yang
Keyword(s):  
Protein Interactions ◽  
White Spot Syndrome Virus ◽  
Envelope Proteins ◽  
White Spot ◽  
Multiprotein Complex ◽  
Yeast Two Hybrid ◽  
White Spot Syndrome ◽  
Self Association ◽  
First Time ◽  
Two Hybrid

ABSTRACT Early events in white spot syndrome virus (WSSV) morphogenesis, particularly the formation of viral membranes, are poorly understood. The major envelope proteins of WSSV are VP28, VP26, VP24, and VP19. Our previous results indicated that VP28 interacts with VP26 and VP24. In the present study, we used coimmunoprecipitation assays and pull-down assays to confirm that the four major proteins in the WSSV envelope can form a multiprotein complex. Yeast two-hybrid assays were also used to test for interactions among the four proteins. In summary, three pairwise protein interactions (VP19-VP28, VP19-VP24, and VP24-VP26) and one self-association (VP24-VP24) were identified for the first time.

Fluorescence Quenching of Aminodiphenylamines with Chloromethanes

2002 ◽  
Vol 67 (8) ◽  
pp. 1154-1164 ◽  
Author(s):  
Nachiappan Radha ◽  
Meenakshisundaram Swaminathan
Keyword(s):  
Charge Transfer ◽  
Fluorescence Quenching ◽  
Ground State ◽  
Rate Constants ◽  
Quenching Mechanism ◽  
Quenching Rate ◽  
Charge Transfer Interaction ◽  
Electronically Excited ◽  
A Charge

The fluorescence quenching of 2-aminodiphenylamine (2ADPA), 4-aminodiphenylamine (4ADPA) and 4,4'-diaminodiphenylamine (DADPA) with tetrachloromethane, chloroform and dichloromethane have been studied in hexane, dioxane, acetonitrile and methanol as solvents. The quenching rate constants for the process have also been obtained by measuring the lifetimes of the fluorophores. The quenching was found to be dynamic in all cases. For 2ADPA and 4ADPA, the quenching rate constants of CCl4 and CHCl3 depend on the viscosity, whereas in the case of CH2Cl2, kq depends on polarity. The quenching rate constants for DADPA with CCl4 are viscosity-dependent but the quenching with CHCl3 and CH2Cl2 depends on the polarity of the solvents. From the results, the quenching mechanism is explained by the formation of a non-emissive complex involving a charge-transfer interaction between the electronically excited fluorophores and ground-state chloromethanes.

scholarly journals Interactome Analysis of KIN (Kin17) Shows New Functions of This Protein

2021 ◽  
Vol 43 (2) ◽  
pp. 767-781
Author(s):  
Vanessa Pinatto Gaspar ◽  
Anelise Cardoso Ramos ◽  
Philippe Cloutier ◽  
José Renato Pattaro Junior ◽  
Francisco Ferreira Duarte Junior ◽  
...  
Keyword(s):  
Dna Replication ◽  
Protein Interactions ◽  
Ribosome Biogenesis ◽  
Cell Metabolism ◽  
Cell Types ◽  
Protein Protein Interactions ◽  
Cellular Mechanisms ◽  
Cancerous Cell ◽  
First Time

KIN (Kin17) protein is overexpressed in a number of cancerous cell lines, and is therefore considered a possible cancer biomarker. It is a well-conserved protein across eukaryotes and is ubiquitously expressed in all cell types studied, suggesting an important role in the maintenance of basic cellular function which is yet to be well determined. Early studies on KIN suggested that this nuclear protein plays a role in cellular mechanisms such as DNA replication and/or repair; however, its association with chromatin depends on its methylation state. In order to provide a better understanding of the cellular role of this protein, we investigated its interactome by proximity-dependent biotin identification coupled to mass spectrometry (BioID-MS), used for identification of protein–protein interactions. Our analyses detected interaction with a novel set of proteins and reinforced previous observations linking KIN to factors involved in RNA processing, notably pre-mRNA splicing and ribosome biogenesis. However, little evidence supports that this protein is directly coupled to DNA replication and/or repair processes, as previously suggested. Furthermore, a novel interaction was observed with PRMT7 (protein arginine methyltransferase 7) and we demonstrated that KIN is modified by this enzyme. This interactome analysis indicates that KIN is associated with several cell metabolism functions, and shows for the first time an association with ribosome biogenesis, suggesting that KIN is likely a moonlight protein.

scholarly journals High-resolution AP-SMALDI MSI as a tool for drug imaging in Schistosoma mansoni

2021 ◽  
Author(s):  
Annika S. Mokosch ◽  
Stefanie Gerbig ◽  
Christoph G. Grevelding ◽  
Simone Haeberlein ◽  
Bernhard Spengler
Keyword(s):  
Schistosoma Mansoni ◽  
Egg Production ◽  
Drug Repurposing ◽  
Cancer Drug ◽  
Ionization Mass ◽  
Parasitic Flatworm ◽  
Paired Female ◽  
First Time ◽  
Organ Tropism

AbstractSchistosoma mansoni is a parasitic flatworm causing schistosomiasis, an infectious disease affecting several hundred million people worldwide. Schistosomes live dioeciously, and upon pairing with the male, the female starts massive egg production, which causes pathology. Praziquantel (PZQ) is the only drug used, but it has an inherent risk of resistance development. Therefore, alternatives are needed. In the context of drug repurposing, the cancer drug imatinib was tested, showing high efficacy against S. mansoni in vitro. Besides the gonads, imatinib mainly affected the integrity of the intestine in males and females. In this study, we investigated the potential uptake and distribution of imatinib in adult schistosomes including its distribution kinetics. To this end, we applied for the first time atmospheric-pressure scanning microprobe matrix-assisted laser desorption/ionization mass spectrometry imaging (AP-SMALDI MSI) for drug imaging in paired S. mansoni. Our results indicate that imatinib was present in the esophagus and intestine of the male as early as 20 min after in vitro exposure, suggesting an oral uptake route. After one hour, the drug was also found inside the paired female. The detection of the main metabolite, N-desmethyl imatinib, indicated metabolization of the drug. Additionally, a marker signal for the female ovary was successfully applied to facilitate further conclusions regarding organ tropism of imatinib. Our results demonstrate that AP-SMALDI MSI is a useful method to study the uptake, tissue distribution, and metabolization of imatinib in S. mansoni. The results suggest using AP-SMALDI MSI also for investigating other antiparasitic compounds and their metabolites in schistosomes and other parasites. Graphical abstract

scholarly journals A Novel Thiosemicarbazide-Based Fluorescent Chemosensor for Hypochlorite in Near-Perfect Aqueous Solution and Zebrafish

Chemosensors ◽  
2021 ◽  
Vol 9 (4) ◽  
pp. 65
Author(s):  
Minji Lee ◽  
Donghwan Choe ◽  
Soyoung Park ◽  
Hyeongjin Kim ◽  
Soomin Jeong ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Reactive Oxygen Species ◽  
Aqueous Solution ◽  
Fluorescence Quenching ◽  
Limit Of Detection ◽  
Fluorescent Sensor ◽  
Ionization Mass ◽  
Oxygen Species ◽  
Marked Selectivity ◽  
Uv Visible

A novel thiosemicarbazide-based fluorescent sensor (AFC) was developed. It was successfully applied to detect hypochlorite (ClO−) with fluorescence quenching in bis-tris buffer. The limit of detection of AFC for ClO− was analyzed to be 58.7 μM. Importantly, AFC could be employed as an efficient and practical fluorescent sensor for ClO− in water sample and zebrafish. Moreover, AFC showed a marked selectivity to ClO− over varied competitive analytes with reactive oxygen species. The detection process of AFC to ClO− was illustrated by UV–visible and fluorescent spectroscopy and electrospray ionization–mass spectrometry (ESI–MS).

scholarly journals Detection of nitroaromatic vapours with diketopyrrolopyrrole thin films: exploring the role of structural order and morphology on thin film properties and fluorescence quenching efficiency

2015 ◽  
Vol 51 (6) ◽  
pp. 1143-1146 ◽  
Author(s):  
Monika Warzecha ◽  
Jesus Calvo-Castro ◽  
Alan R. Kennedy ◽  
Alisdair N. Macpherson ◽  
Kenneth Shankland ◽  
...  
Keyword(s):  
Thin Film ◽  
Thin Films ◽  
Fluorescence Quenching ◽  
Optical Detection ◽  
Film Properties ◽  
Structural Order ◽  
Quenching Efficiency ◽  
First Time

Sensitive optical detection of nitroaromatic vapours with diketopyrrolopyrrole thin films is reported for the first time.

Heteroarray of cobalt(II)tetrasulfophthalocyanine and cobalt(II) tetrakis(N-methyl-4-pyridyl)porphyrin: synthesis, isolation and electronic properties

2012 ◽  
Vol 16 (02) ◽  
pp. 244-249 ◽  
Author(s):  
Thiago T. Tasso ◽  
Wania C. Moreira
Keyword(s):  
Electronic Properties ◽  
Spectroscopic Characterization ◽  
Charge Transfer Process ◽  
Isolation And Purification ◽  
Electronic Densities ◽  
Porphyrin Synthesis ◽  
A Charge ◽  
Job's Method ◽  
First Time

Porphyrin and phthalocyanine macrocycles with ionic substituents can form mixed assemblies with interesting electronic properties for potential application on the development of new devices. This paper reports the synthesis, isolation and purification of heteroaggregate formed by cobalt(II) 4,4′,4″,4‴-tetrasulfophthalocyanine (CoTsPc) and cobalt(II) tetrakis(N-methyl-4-pyridyl)porphyrin (CoTMPyP), followed by its spectroscopic characterization. Spectroscopic titration, performed with a CoTsPc water/acetone solution, allowed the use of Job's method for determination of the heteroaggregates stoichiometry. The Job's plot revealed the formation of only one predominant heterocomplex in solution, containing two CoTsPc molecules in terminal positions and a central CoTMPyP one. For the first time, a method for isolation and purification of a mixed ionic array has been reported in the literature. The triad's electronic spectrum is quite different from the sum of the macrocycles isolated spectra, due to the overlap of their electronic densities and a charge transfer process between CoTsPc and CoTMPyP.

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