210107-35-0Relevant academic research and scientific papers
Photoaffinity palladium reagents for capture of protein-protein interactions
Zheng, Qizhen,Pang, Zhengyuan,Liu, Jingwei,Zhou, Yi,Sun, Yang,Yin, Zheng,Lou, Zhiyong
, p. 6369 - 6373 (2019)
Protein-protein interactions (PPIs) are indispensable in almost all cellular processes. Probing of complex PPIs provides new insights into the biological system of interest and paves the way for the development of therapeutics. Herein, we report a strategy for the capture of protein-protein interactions using photoaffinity palladium reagents. First, the palladium-mediated reagent site specifically transferred a photoaffinity modified aryl group to the designated cysteine residue. Next, the photoaffinity group was activated by UV radiation to trap the proximal protein residue for the formation of a crosslink. This strategy was used to capture the PYL-ABA-PP2C interaction, which is at the core of the abscisic acid (ABA) signalling pathway. Our results indicated that this palladium-mediated strategy can serve as an alternative for incorporating an increasing number of diverse substrates for protein crosslinking through cysteine modifications and can be explored for use in mapping protein-peptide or protein-protein interaction surfaces and in trapping potential interacting partners.
Allyl m -trifluoromethyldiazirine mephobarbital: An unusually potent enantioselective and photoreactive barbiturate general anesthetic
Savechenkov, Pavel Y.,Zhang, Xi,Chiara, David C.,Stewart, Deirdre S.,Ge, Rile,Zhou, Xiaojuan,Raines, Douglas E.,Cohen, Jonathan B.,Forman, Stuart A.,Miller, Keith W.,Bruzik, Karol S.
experimental part, p. 6554 - 6565 (2012/09/21)
We synthesized 5-allyl-1-methyl-5-(m-trifluoromethyl-diazirynylphenyl) barbituric acid (14), a trifluoromethyldiazirine-containing derivative of general anesthetic mephobarbital, separated the racemic mixture into enantiomers by chiral chromatography, and determined the configuration of the (+)-enantiomer as S by X-ray crystallography. Additionally, we obtained the 3H-labeled ligand with high specific radioactivity. R-(-)-14 is an order of magnitude more potent than the most potent clinically used barbiturate, thiopental, and its general anesthetic EC50 approaches those for propofol and etomidate, whereas S-(+)-14 is 10-fold less potent. Furthermore, at concentrations close to its anesthetic potency, R-(-)-14 both potentiated GABA-induced currents and increased the affinity for the agonist muscimol in human α1β2/3γ2L GABAA receptors. Finally, R-(-)-14 was found to be an exceptionally efficient photolabeling reagent, incorporating into both α1 and β3 subunits of human α1β3 GABA A receptors. These results indicate R-(-)-14 is a functional general anesthetic that is well-suited for identifying barbiturate binding sites on Cys-loop receptors.
Stereospecific synthesis of a carbene-generating angiotensin II analogue for comparative photoaffinity labeling: Improved incorporation and absence of methionine selectivity
Pillion, Dany,Dera?t, Maud,Holleran, Brian J.,Escher, Emanuel
, p. 2200 - 2209 (2007/10/03)
A stereospecific convergent synthesis of N-[(9-fluorenyl)methoxycarbonyl]- p-[3-(trifluoromethyl)-3H-diazirin-3-yl]-L-phenylalanine (Fmoc-12, Fmoc-Tdf) and its incorporation into the C-terminal position of the angiotensin II (AngII) peptide to form 125I[Sar1,Tdf8]AngII ( 125I-13) is presented. This amino acid photoprobe is a highly reactive carbene-generating diazirine phenylalanine derivative that can be used for photoaffinity labeling. Using model receptors, we compared the reactivity and the Met selectivity of 12 to that of the widely used and reputedly Met-selectivep-benzoyl-L-phenylalanine (Bpa) photoprobe. Wild-type and mutant AngII type 2 receptors, a G protein-coupled receptors, were photolabeled with 125I-13 as well as with 125I-[Sar1,Bpa 8]AngII (125I-14), and the respective incorporation yields were assessed. The carbene-generating 12 was more reactive toward inert residues and was not Met-selective compared to the biradical ketone-generating Bpa, allowing for more precise determination of ligand contact points in peptidergic receptors.
Synthesis of a new photo-cross-linking nucleoside analogue containing an aryl(trifluoromethyl)diazirine group: Application for EcoRII and MvaI restriction-modification enzymes
Topin, Andrey N.,Gritsenko, Oxana M.,Brevnov, Maxim G.,Gromova, Elisaveta S.,Korshunova, Galina A.
, p. 1163 - 1175 (2007/10/03)
A new photo-cross-linking dU analog, 5-[4-(3-(trifluoromethyl)-3H- diazirin-3-yl)phenyl]-2'-deoxyuridine, was synthesized and incorporated into the recognition site of EcoRII and MvaI restriction-modification enzymes. The resulting base-modified 14-mer su
