For full access to this pdf, sign in to an existing account, or purchase an annual subscription. Staudinger ligation technology spotlights sigmaaldrich. Center for new directions in organic synthesis, department of chemistry, university of. A chemical transformation that permits the selective formation of covalent adducts among richly functionalized biopolymers within a cellular context is presented. Traceless staudinger ligation for the synthesis of peptides and proteins in water inventors ronald raines, matthew soellner, annie tam since its founding in 1925 as the patenting and licensing organization for the university of wisconsinmadison, warf has been working with business and industry to transform university research into products. Following this new methodology, the phosphine oxide10 that remains in the nontraceless procedure is excised from the target molecule during. The staudinger ligationa gift to chemical biology request pdf. Staudinger bertozzi ligation between compound x or xii and an azidecontaining biomolecule yields, respectively, an 20 or 24 a linker between the biomolecule and the fluorophore. Catalytic staudingervilarrasa reaction for the direct.
Lightactivated staudingerbertozzi ligation within living animals. Pdf the traceless staudinger ligation for indirect 18f. The traceless staudinger ligation for indirect 18f. Staudinger reactions for selective functionalization of. Although the subsequent staudinger ligation was performed under argon, only low conversion could be observed for these types of phosphines 37 % and. It was not until recently, however, that bertozzi and co. Catalystfree chemoselective dna conjugation by the. The staudinger ligation of phosphinesubstituted thioesters with 18ffluoroethylazide has been successfully applied to access 18flabelled molecules in radiochemical yields superior to 95%. As the label we used either coumarin4acetic acids 1ac or ferrocene derivative 17, which were synthesised in our department as part of studies into new ligands. A new double staudinger linker has been specifically developed to.
Chemoselective ligation refers to the use of mutually specific reactive chemical groups to accomplish molecular conjugation. Hermann staudinger 23 march 1881 8 september 1965 was a german organic chemist who demonstrated the existence of macromolecules, which he characterized as polymers. In combination with nonnatural protein translation the staudinger. The traceless staudinger ligation has been applied to the assembly of a protein from constituent peptides nilsson et al. A highyield, chemoselective, and mild synthetic method. Our efforts to develop a scalable and divergent synthesis of cyclic dinucleotide analog precursors have resulted in 1 an orthogonally protected diamino carbohydrate as well as 2 the novel application of the staudinger ligation to provide mediumsized macrocycles featuring carbamate or urea linkages. A novel application of the staudinger ligation to access. The rapid advancement of bioorthogonal click chemistry in the past decade has enabled the study and precise manipulation of biological processes within living organisms. In 1880 there were 17 staudinger families living in new york. Pdf staudinger ligation towards cyclodextrin dimers in aqueous. The staudinger reaction is the reaction between an azide and phosphine, which form a reactive azaylide as an intermediate that is hydrolyzed spontaneously to give a primary amine and a phosphine oxide 89,90. These attributes endow the staudinger ligation with broad utility. Synthesis of coumarin or ferrocene labeled nucleosides via.
The staudinger ligation is a bioorthogonal reaction between azides and phosphines that requires no catalyst to proceed, allowing for mild reaction conditions. Selective chemical reactions enacted within a cellular environment can be powerful tools for elucidating biological processes or engineering novel interactions. Department of chemistry, university of konstanz, 78457, konstanz, germany. A new technique based on the staudinger ligation has enabled researchers to conduct a broad analysis of cellular targets of protein farnesylation, with potential implications for future proteomic. Staudinger ligation as a method for bioconjugation deepdyve. The staudinger family name was found in the usa, and canada between 1880 and 1920. Cd dimers in good yields under mild conditions in aqueousorganic media. Watersoluble phosphinothiols for traceless staudinger. Metabolic labeling of glycans with azido sugars and. Staudinger equation definition of staudinger equation by. View enhanced pdf access article on wiley online library html view download pdf for offline viewing. Azides may be converted to amines by hydrogenation, but another possibility is the staudinger reaction, which is a very mild azide reduction. Here, this restriction is overcome with a new chemical ligation method in which a phosphinobenzenethiol is used to link a thioester and azide. Staudinger ligation reaction chemistry thermo fisher.
Staudinger equation definition is an equation for determining the molecular weight of polymeric materials that utilizes the viscosity of solutions of the polymer at definite concentrations. In the course of their studies on the metabolic engineering of cell surfaces they designed a phosphine with an ester moiety as an intramolecular electrophilic trap. An fmoc compatible, o to s shiftmediated procedure for the preparation of cterminal thioester peptides. Staudinger reaction an overview sciencedirect topics.
Catalystfree chemoselective dna conjugation by the staudinger ligation and diels alder cycloaddition reaction. The present work demonstrates the application of the staudinger ligation reaction for the first time to form. The staudinger ligation, an amide bond forming reaction between an azide and a phosphine containing ester, was developed by saxon et al, a traceless staudinger ligation for chemoselective synthesis of amide bonds, organic letters, american chemical society, vol. Hydrazone ligation oxime ligation thiazolidine ligation 5 m. This was about 39% of all the recorded staudingers in the usa. For this work he received the 1953 nobel prize in chemistry. Catalytic staudingervilarrasa reaction for the direct ligation of carboxylic acids and azides doi. This reaction is frequently used for the study of lipids, steroids, and cofactors.
Cell surface engineering by a modified staudinger reaction. His work laid the foundation for the great expansion of the plastics industry later in the 20th century. This method is limited, however, by an absolute requirement for a cysteine residue at the ligation juncture. Staudinger related reactions, such as staudinger reduction, staudinger ligation, and traceless staudinger ligation comprise a powerful family of techniques enabling preparation of a wide range of polysaccharide derivatives with excellent chemoselectivity and the potential for excellent regioselectivity when combined with other methods. This method for the selective formation of an amide bond, which does not require the orthogonal protection of distal functional groups, should find general utility in synthetic and biological chemistry. The capability to induce fast and selective chemical reactions between two exogenous complementary moieties in living systems, with negligible perturbation to their native activities, have rendered bio. Esiptbased highly selective fluorescent probe for organic azides. A pro linebased phosphine template for staudinger ligation. Mechanistic investigation of the staudinger ligation journal of the. Watersoluble phosphinothiols for traceless staudinger ligation and integration with expressed protein ligation.
Triple bioorthogonal ligation strategy for simultaneous. Staudinger ligation towards cyclodextrin dimers in aqueous. Files are available under licenses specified on their description page. Triple bioorthogonal ligation strategy for simultaneous labeling of multiple enzymatic activities article in angewandte chemie international edition 5118. We used intermolecular staudinger ligation for the preparation of fluorescent and electrochemically labeled nucleosides. Synthesis of alexa647phosphine 20 a a and alexa488phosphine 24 a b.
Given these limitations, it appears that at least five different ligation products are formed based on the western blot analysis. Staudinger chemistry, based on the specific crosslinking reaction between azide and phosphinelabeled molecules, is one of the more promising and versatile examples of chemoselective strategies for applications such as metabolic labeling. Thus, staudinger ligation could be the method of choice for linking cds exhibiting i ease of. Origin of problems related to staudinger reduction in.
The staudinger reaction is a chemical reaction of an azide with a phosphine or phosphite. Other staudinger ligation induced macrocyclizations have been published previously by maarseveen and coworkers, who successfully used the raines ligation reagent for the synthesis of a series of mediumsized lactams. Staudinger studied chemistry at the universities of. Sitespecific and chemoselective labeling of dna is still a difficult task. Hermann staudinger, german chemist who won the 1953 nobel prize for chemistry for demonstrating that polymers are longchain molecules. Protein engineering with the traceless staudinger ligation. Coulombic effects on the traceless staudinger ligation in. The most staudinger families were found in the usa in 1920. As there are a variety of methods for preparing azides readily, the staudinger reaction makes it possible to use n 3 as an nh 2 synthon. Here we report a novel modification of our previously reported staudinger ligation that generates an amide bond from an azide and a specifically functionalized phosphine.
Pdf hermann staudingers pioneering theories on the polymer structures of fibers and plastics and his later research on biological macromolecules formed the basis for countless modern developments in the fields of materials science and biosciences and supported the rapid growth of the. To optimize the conditions for staudinger ligation of mdhfr3, a range of temperatures, triarylphosphine. Bioorthogonal click chemistry for in vivo bioimaging. Catalytic staudingervilarrasa reaction for the direct ligation. Staudinger ligation as a method for bioconjugation staudinger ligation as a method for bioconjugation van berkel, sander s van eldijk, mark b van hest, jan c. Synthetic glycomimetics, derived by mimicking complex glycoligands on a variety of scaffolds, have become important tools for studying the role of carbohydrates in chemistry. Snitrosothiols rsnos represent an important class of. Of interest in chemical biology research, the staudinger ligation is a modification of the classical staudinger reaction in which an electrophilic trap usually a methyl ester is placed on the triaryl phosphine. Staudinger ligation tfa, 95 % acetonitrile in water b and uv detection at 220. Metabolic labeling of glycans with azido sugars and subsequent glycanprofiling and visualization via staudinger ligation.
Highyielding staudinger ligation of a phosphinothioester and. A novel esipt type fluorescent probe was developed for detection of organic azides through staudinger ligation to release the masked fluorophore. A traceless staudinger ligation for the chemoselective. All structured data from the file and property namespaces is available under the creative commons cc0 license. The mechanism begins with attack of the phosphorus on the far nitrogen of. Traceless staudinger ligation for the synthesis of. Staudinger ligation as a method for bioconjugation van. Based on staudinger ligation, a novel esiptbased fluorescent. To assess our reasoning on coulombic effects on the traceless staudinger ligation in water, we used phosphinothiols 24 to effect a model dipeptide coupling with clabeled azidoglycine 27 10. Getting a handle on protein farnesylation nature methods. Dna conjugation by staudinger ligation nucleic acids. Phosphinothioesters 2326 were synthesized by the reaction of the respective phosphinothiol 14 with acglyopfp in the presence of diea.
New york had the highest population of staudinger families in 1880. Incorporation of azides into recombinant proteins for chemoselective modification by the staudinger ligation kristi l. Although the previously described methods for staudinger ligations work well even in biological environments, a modification forming a native amide bond. The investigation of heteromultivalent interactions of complex glycoligands and proteins is critical for understanding important biological processes and developing carbohydratebased pharmaceutics. Azidespecific labeling of biomolecules by staudinger. The nsr is much faster with tetrafluorinated aromatic azide than that the staudingerbertozzi or spaac ligation with alkyl azide, as revealed by hplc analysis and fluorescence kinetics.
The staudinger reaction is an organic reaction used to convert an organic azide to a primary amine using a pr 3 compound such as triphenylphosphine and water. Originally reported by staudinger and meyer, azides react readily with triarylphosphines to form the corresponding iminophophoranes. The latter reaction is commonly referred to as the staudinger ligation. The technique of native chemical ligation enables the total chemical synthesis of proteins. A pro linebased phosphine template for staudinger ligation article in organic letters 1417. In 1919 the german chemist hermann staudinger was the first to describe the reaction between an azide and a phosphine. Incorporation of azides into recombinant proteins for. Bertozzi and coworkers demonstrated the staudinger ligation of an azide sianaz and a functionalized phosphine phosflag in living mice. Coumarins as fluorescent probes or labels have extensive and diverse applications.