Single-armed phenylsulfonated pyridine derivative of DOTA is rigid and stable paramagnetic tag in protein analysis

2016 ◽  
Vol 52 (77) ◽  
pp. 11535-11538 ◽  
Author(s):  
Feng Yang ◽  
Xiao Wang ◽  
Bin-Bin Pan ◽  
Xun-Cheng Su
Keyword(s):  
Long Range ◽  
Protein Analysis ◽  
Pyridine Derivative ◽  
Pyridine Derivatives ◽  
Site Specific ◽  
Protein Conjugates ◽  
Paramagnetic Tag ◽  
Structural Restraints ◽  
Respective Protein ◽  
Specific Labelling

Single-armed DOTA-like phenylsulfonated pyridine derivatives are rigid and stable paramagnetic tags for site-specific labelling of proteins. The respective protein conjugates yield valuable long-range structural restraints for proteins.

Multinuclear NMR spectra of [Pt(L)Cl3]− (L = pyridine derivatives) complexes and crystal structure of trans-Pt(2,6-di(hydroxymethyl)pyridine)2Cl2•2H2O

1996 ◽  
Vol 74 (11) ◽  
pp. 2121-2130 ◽  
Author(s):  
Fernande D. , ◽  
Corinne Bensimon ◽  
André L. Beauchamp
Keyword(s):  
Crystal Structure ◽  
Pyridine Ring ◽  
Chemical Shifts ◽  
Bond Distance ◽  
Coupling Constants ◽  
Mirror Plane ◽  
Pyridine Derivative ◽  
Pyridine Derivatives ◽  
Pyridine Ligand ◽  
Twofold Axis

Complexes of the type [Pt(L)Cl3]− (L = pyridine derivative) were synthesized and studied by 13C and 195Pt NMR spectroscopies. The 195Pt signals were observed between −1720 and −1897 ppm. No correlation between the δ(Pt) and the pKa of the protonated pyridine derivatives was found. The chemical shifts vary with the substituents on the pyridine ligand. Compounds with substituents in ortho positions were observed at lower fields, except for complexes containing hydroxy or amine groups. The latter compounds were observed at higher fields, close to the signals of the Pt-unsubstituted pyridine compound. These results were explained in terms of the solvent effect. The chemical shifts δ(C) and the coupling constants J(13C–195Pt) were measured and the results interpreted with a view of obtaining information on the nature of the Pt—N bond. The possibility of π-bonding between platinum and the pyridine ligand is examined. The conformation of the pyridine ring in relation to the platinum plane and the energies of the rotation barriers around the Pt—N bond in these types of platinum(II) complexes are briefly discussed. The crystal structure of trans-Pt(2,6-(HOCH2)2py)2Cl2•2H2O was determined by X-ray diffraction. The compound is monoclinic, C2/m, a = 7.022(6), b = 15.646(13), c = 8.344(10) Å, β = 93.35(8)°, Z = 2, R = 0.037. The platinum atom is located at the junction of the twofold axis and the mirror plane, the N atoms and the para-C atom of the pyridine ring are situated on the twofold axis, and the chloride ligands are on the mirror plane. The compound crystallizes with molecules of water, which are H-bonded to the hydroxy groups. The Pt—Cl bond distance is 2.306(2) Å, and that of the Pt—N bond is 2.041 (6) Å. The dihedral angle between the platinum and the pyridine planes is 79.8°. Key words: platinum, pyridine derivatives, NMR, crystal structure.

Site-Specific Labelling of Native Mammalian Proteins for Single-Molecule FRET Measurements

ChemBioChem ◽  
2018 ◽  
Vol 19 (8) ◽  
pp. 780-783 ◽  
Author(s):  
Alexander Gust ◽  
Leonhard Jakob ◽  
Daniela M. Zeitler ◽  
Astrid Bruckmann ◽  
Kevin Kramm ◽  
...  
Keyword(s):  
Single Molecule ◽  
Site Specific ◽  
Single Molecule Fret ◽  
Mammalian Proteins ◽  
Specific Labelling

scholarly journals Bioorthogonal protein-DNA conjugation methods for force spectroscopy

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Marie Synakewicz ◽  
Daniela Bauer ◽  
Matthias Rief ◽  
Laura S. Itzhaki
Keyword(s):  
Single Molecule ◽  
Unnatural Amino Acids ◽  
Force Spectroscopy ◽  
Diels Alder ◽  
Cysteine Residues ◽  
Single Molecule Force Spectroscopy ◽  
Site Specific ◽  
Inverse Electron Demand ◽  
Protein Conjugates ◽  
Electron Demand

Abstract Accurate and stable site-specific attachment of DNA molecules to proteins is a requirement for many single-molecule force spectroscopy techniques. The most commonly used method still relies on maleimide chemistry involving cysteine residues in the protein of interest. Studies have consequently often focused on model proteins that either have no cysteines or with a small number of cysteines that can be deleted so that cysteines can then be introduced at specific sites. However, many proteins, especially in eukaryotes, contain too many cysteine residues to be amenable to this strategy, and therefore there is tremendous need for new and broadly applicable approaches to site-specific conjugation. Here we present bioorthogonal approaches for making DNA-protein conjugates required in force spectroscopy experiments. Unnatural amino acids are introduced site-specifically and conjugated to DNA oligos bearing the respective modifications to undergo either strain-promoted azidealkyne cycloaddition (SPAAC) or inverse-electron-demand Diels-Alder (IE-DA) reactions. We furthermore show that SPAAC is compatible with a previously published peptide-based attachment approach. By expanding the available toolkit to tag-free methods based on bioorthogonal reactions, we hope to enable researchers to interrogate the mechanics of a much broader range of proteins than is currently possible.

Synthesis of novel maleimide derivatives for “site-specific labelling”

2010 ◽  
Vol 37 (6) ◽  
pp. 689
Author(s):  
Verena Nagel ◽  
Carsten Burchardt ◽  
Patrick J. Riss ◽  
Frank Rösch
Keyword(s):  
Site Specific ◽  
Specific Labelling

Pyrazolo[3,4-b]pyridine in heterocyclic synthesis: synthesis of new pyrazolo[3,4-b]pyridines, imidazo[1',2':1,5]pyrazolo[3,4-b]pyridines, and pyrido[2',3':3,4]pyrazolo[1,5-a]pyrimidines

2007 ◽  
Vol 85 (9) ◽  
pp. 592-599 ◽  
Author(s):  
Mohamed A.M Gad-Elkareem ◽  
Azza M Abdel-Fattah ◽  
Mohamed A.A Elneairy
Keyword(s):  
Pyridine Derivative ◽  
Pyridine Derivatives ◽  
Heterocyclic Synthesis ◽  
Acetyl Acetone ◽  
Active Methylene

Pyrazolo[3,4-b]pyridine derivatives 7 and 9 were synthesized via the reaction of 3-amino-1H-pyrazolo-[3,4-b]pyridine derivative 2 with ω-bromoacetophenones. Reaction of 7 and 9 with Ac2O afforded the imidazo[1',2':1,5]py razolo[3,4-b]pyridine derivative 8 and pyrazolo[3,4-b]pyridine derivative 10, respectively. Reaction of 2 with chloroacetonitrile followed by DMF-DMA gave imidazo[1',2':1,5]pyrazolo[3,4-b]pyridines 4 and 5, respectively. Acetyl acetone and 1,1-dicyano-2,2-dimethylthioethene were reacted with 2 to afford the pyrido[2',3':3,4]pyrazolo-[1,5-a]-pyrimidines 11 and 14, respectively. Also, 2 reacted with DMF-DMA to yield the formamidine 15, which in turn, reacted with active methylene reagents, yielding the corresponding pyrido[2',3':3,4]pyrazolo[1,5-a]pyrimidines 18 and 23a-23d.Key words: 1H-pyrazolo[3,4-b]pyridines, imidazo[1',2':1,5]pyrazolo[3,4-b]pyridines, pyrido[2',3':3,4]pyrazolo[1,5-a]pyrimidines.

Thiol–ene reaction: a versatile tool in site-specific labelling of proteins with chemically inert tags for paramagnetic NMR

2012 ◽  
Vol 48 (21) ◽  
pp. 2704 ◽  
Author(s):  
Qing-Feng Li ◽  
Yin Yang ◽  
Ansis Maleckis ◽  
Gottfried Otting ◽  
Xun-Cheng Su
Keyword(s):  
Paramagnetic Nmr ◽  
Site Specific ◽  
Ene Reaction ◽  
Versatile Tool ◽  
Specific Labelling

Reinvestigation of the Reaction of Arylidene Malononitrile with Cyanothioacetamide: New Approach for the Synthesis of Pyridine Derivatives

1992 ◽  
Vol 47 (10) ◽  
pp. 1438-1440 ◽  
Author(s):  
A. A. Geies ◽  
A. M. Kamal El-Dean ◽  
M. I. Abdel Monem
Keyword(s):  
Diethyl Malonate ◽  
Pyridine Derivative ◽  
Pyridine Derivatives ◽  
New Approach

Arylidene malononitrile reacted with cyanothioacetamide under basic condition to afford the pyridine derivative (2). The same product was obtained from the reaction of arylidene ethylcyanoacetate, arylidene diethyl malonate, arylidene pyrazolinone with cyanothioacetamide under the same condition. A resonable mechanism for this behaviour was suggested.

Site-Specific Labelling of Proteins with a Rigid Lanthanide-Binding Tag

ChemBioChem ◽  
2006 ◽  
Vol 7 (10) ◽  
pp. 1599-1604 ◽  
Author(s):  
Xun-Cheng Su ◽  
Thomas Huber ◽  
Nicholas E. Dixon ◽  
Gottfried Otting
Keyword(s):  
Site Specific ◽  
Specific Labelling

scholarly journals The Novel Monocomponent FAD-dependent Monooxygenase HpaM Catalyzes the 2-Decarboxylative Hydroxylation of 5-Hydroxypicolinic Acid inAlcaligenes faecalisJQ135

2017 ◽  
Author(s):  
Jiguo Qiu ◽  
Bin Liu ◽  
Lingling Zhao ◽  
Yanting Zhang ◽  
Dan Cheng ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Microbial Degradation ◽  
Pyridine Ring ◽  
Degradation Process ◽  
Alcaligenes Faecalis ◽  
Pyridine Derivative ◽  
Pyridine Derivatives ◽  
Monooxygenase Activity ◽  
E Coli ◽  
Transformation Mechanisms

Abstract5-hydroxypicolinic acid (5HPA) is a natural pyridine derivative that can be microbially degraded. However, the physiological, biochemical, and genetic foundation of the microbial catabolism of 5HPA remains unknown. In this study, a gene clusterhpa(which is involved in degradation of 5HPA inAlcaligenes faecalisJQ135) was cloned and HpaM was identified as a novel monocomponent FAD-dependent monooxygenase. HpaM shared a sequence only 31% similarity with the most related protein 6-hydroxynicotinate 3-monooxygenase (NicC) ofPseudomonas putidaKT2440.hpaMwas heterologously expressed inE. coliBL21(DE3), and the recombinant HpaM was purified via Ni-affinity chromatography. HpaM catalyzed the 2-decarboxylative hydroxylation of 5-HPA, thus generating 2,5-dihydroxypyridine (2,5-DPH). Monooxygenase activity was only detected in the presence of FAD and NADH, but not of FMN and NADPH. The apparentKmvalues of HpaM toward 5HPA and NADH were 45.4 μ and 37.8 μ, respectively. Results of gene deletion and complementation showed thathpaMwas essential for 5HPA degradation inAlcaligenes faecalisJQ135.ImportancePyridine derivatives are ubiquitous in nature and important chemical materials that are currently widely used in agriculture, pharmaceutical, and chemical industries. Thus, the microbial degradation and transformation mechanisms of pyridine derivatives received considerable attention. Decarboxylative hydroxylation was an important degradation process in pyridine derivatives, and previously reported decarboxylative hydroxylations happened in the C3 of the pyridine ring. In this study, we cloned the gene clusterhpa, which is responsible for 5HPA degradation inAlcaligenes faecalisJQ135, thus identifying a novel monocomponent FAD-dependent monooxygenase HpaM. Unlike 3-decarboxylative monooxygenases, HpaM catalyzed decarboxylative hydroxylation in the C2 of the pyridine ring in 5-hydroxypicolinic acid. These findings deepen our understanding of the molecular mechanism of microbial degradation of pyridine derivatives. Furthermore, HpaM offers potential for applications to transform useful pyridine derivatives.

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