Bjoc staudinger ligation towards cyclodextrin dimers in aqueous. If the reaction were to proceed similarly within an intact cell, functionalities such as spin labels, fluorophores, and radiolabels may be appended to a phosphine and delivered to an azidelabeled. Chemoselective ligation refers to the use of mutually specific reactive chemical groups to accomplish molecular conjugation. Catalystfree chemoselective dna conjugation by the. Staudinger ligation as a method for bioconjugation. Ligations mediated by phosphinoalcohols had lower rate constants or less chemoselectivity.
It was not until recently, however, that bertozzi and coworkers recognized the potential of this reaction as a method for bioconjugation and transformed it into the socalled staudinger ligation. Azidopeptides6 and azidoproteins19 are readily attainable, and the reactivity of the azido group is. The solubility of the phosphine is a critical parameter that must be consider for successful bioconjugation of 24. While the staudinger reaction has first been described a hundred years ago in 1919, the ligation reaction became one of the most important and efficient bioconjugation techniques in the 1990s and this century. Download chemical ligation book pdf epub mobi tuebl and. Staudinger reaction an overview sciencedirect topics. Sitespecific protein immobilization by staudinger ligation. Azides may be converted to amines by hydrogenation, but another possibility is the staudinger reaction, which is a very mild azide reduction. So far, staudinger ligation has been applied in diverse fields such as protein immobilization, solidphase peptide synthesis, stereoselective nglycosylation, and the synthesis of homogenous glycoproteins. A comprehensive listing of available organic azides and functionalized alkynes rounds off this issue of chemfiles with valuable building blocks for click chemistry or staudinger ligation. Download chemical ligation book pdf epub mobi tuebl and read. It holds the crucial characteristics for bioorthogonal chemistry.
Thus, first we developed the synthesis of the nucleoside triphosphate 1,a29deoxyadenosine analogue scheme 1a. Following this new methodology, the phosphine oxide10 that remains in the nontraceless procedure is. The resulting photochromes show stable and reversible switching behavior in water, with a photostationary state pss of up to 95. The staudinger ligation chemoselective and bioorthogonal. Files are available under licenses specified on their description page. A nucleoside triphosphate for sitespecific labelling of dna. Staudinger ligation for glycoprotein synthesis creative biolabs. 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. Mar 17, 2000 selective chemical reactions enacted within a cellular environment can be powerful tools for elucidating biological processes or engineering novel interactions.
In addition to a known linker, methyl 2diphenylphosphanylterephthalate, a doubly active linker was specifically developed that enabled connection of two. An intramolecular traceless staudinger ligation was employed to synthesize cyclic peptides. The intermediate, a phosphaza ylide 1, scheme 1a is readily formed with loss of nitrogen gas, while subsequent hydrolysis yields the corresponding amine and phosphine oxide. The traceless staudinger ligation for indirect 18f. The final amide bond formation was induced by deprotection of the azidopeptide phosphinothioesters. A novel family of azobenzenes containing residues needed for aqueous staudinger bertozzi ligation to azides was designed see scheme. A putative strategy for the assembly of proteins is depicted in fig. Staudinger reaction major reference works wiley online. Staudinger ligation reaction chemistry thermo fisher.
Liu, 2006 staudinger ligation for glycoprotein synthesis at creative biolabs. Chemoselective crosslinking and functionalization of. A variant of this reaction is the staudinger ligation, a bioorthogonal reaction that has become an important tool of. Dna conjugation by staudinger ligation nucleic acids. A ligation modeled after the staudinger reaction forms an amide bond by. The staudinger reduction is conducted in two steps. Staudinger ligation with a suitably functionalized phosphine.
The staudinger ligation mediated by diphenylphosphinomethanethiol proceeded with a secondorder rate constant 7. Since it has been shown that 7modified 7deaza2 9deoxyadenosine derivatives are. Incorporation of azides into recombinant proteins for. The traceless staudinger ligation enables the formation of an amide bond between a phosphinothioester or phosphinoester and an azide without the incorporation of residual atoms. In 1919, hermann staudinger, future nobel laureate and forefather of polymer chemistry, described a roomtemperature reaction that could be. Jan 08, 2002 the selectivity of the staudinger ligation even within a complex cell lysate mixture suggests the exciting prospect of intracellular applications. In 1919 the german chemist hermann staudinger was the first to describe the reaction between an azide and a phosphine.
The reaction proceeds with high stereoselectivity, and a variety of glycosyl azides can function as substrates. Oct 28, 2020 in 1920, at age 39 and the peak of his professional carrier, one of the most distinguished organic chemists of his generation, hermann staudinger, claimed that natural and synthetic polymers are covalent macromolecules rather than colloidal aggregates. For this work he received the 1953 nobel prize in chemistry. Theoretical geometries of intermolecular dimers of 4 and of the complex 4ada are also shown. The supporting information provides full experimental procedures and detailed analytical data for the synthesis of all compounds. A nucleoside triphosphate for sitespecific labelling of. The onepot nonhydrolysis staudinger reaction and staudinger. Based on the tandem nsr and staudinger bertozzi ligation, we prepare a molecular beacon 7 from 1 in onepot synthesis with a recovery yield of 32%. The staudinger ligation can be used to couple a peptide with a cterminal phosphinothioester to another with an nterminal aazido group to form a single peptide that contains no residual atoms. The staudinger ligation has been developed as a versatile bioorthogonal conjugation tool for a wide variety of biological applications both in vitro and in vivo. The staudinger ligation proofed to be a very chemoselective reaction by numerous in vivo studies. Nglycosyl amides are needed to generate glycopeptides and glycoproteins.
When a faster spaac ligation is used instead of the staudinger bertozzi ligation, compound 8 is prepared in the tandem nsr and spaac reactions with a recovery yield of 59%. Staudinger ligation the staudinger ligation is a reaction developed by the bertozzi group in 2000 that is based on the classic staudinger reaction of azides with triarylphosphines. All structured data from the file and property namespaces is available under the creative commons cc0 license. Selective traceless staudinger ligation for glycoprotein synthesis reveals scope and limitations dr. Jun 09, 2000 engrammic, leaving omercaptobenzylamine in the ligation product. A novel application of the staudinger ligation to access neutral cyclic dinucleotide analog precursors via a divergent method m. Sep 15, 2010 staudinger reaction staudinger ligation, staudinger reduction first published. Subsequent hydrolysis of 9 yields free amide11 and phosphine. Sep 12, 2011 moreover, the staudinger ligation has been used as a synthetic method to construct glycopeptides, microarrays, and functional biopolymers. This idea initiated chain reactions supporting and dismissing the covalent macromolecular concept.
Berkeley, ca no, the staudinger ligation isnt the latest robert ludlum thriller. Base modified 2deoxyadenosine and thymidine 5 triphosphates linked to an azido moiety by different linkers were used to synthesize single azidofunctionalized dna enzymatically by dna polymerases. 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. The staudinger ligation of phosphinesubstituted thioesters with 18ffluoroethylazide has been successfully applied to access 18flabelled molecules in radiochemical yields superior to 95%. Staudinger ligation towards cyclodextrin dimers in aqueousorganic media. 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. Here, we report that the staudinger ligation can be used to form glycosylated asparagine derivatives. For sitespecific labelling of dna the staudinger ligation is an interesting candidate and was further investigated in this work. Presenting a wide array of information on chemical ligation one of the more powerful tools for protein and peptide synthesis this book helps readers understand key methodologies and.
Cd dimers have been prepared using the bioorthogonal staudinger ligation for the first time. Reaction mechanism and kinetics of the traceless staudinger. Due to this adaptability, this robust ligation type is a perfect candidate with a high potential for various applications in the field of radiopharmacy for the labeling of biomolecules under mild conditions. Abstract in 1919 the german chemist hermann staudinger was the first to describe the reaction between an azide and a phosphine. As there are a variety of methods for preparing azides readily, the staudinger reaction makes it possible to use n 3 as an. Here we report a smallmolecule switch that can be used to. The staudinger bertozzi ligation has also been a powerful tool for imaging and investigating biomolecules including glycans, lipids, proteins and dna. From its introduction onwards, the staudinger ligation and the highly. 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. The staudinger reaction is a classical method for the preparation of amines from phosphines and azides. It was not until recently, however, that bertozzi and co. Here, we present the traceless staudinger ligation as a strategy for the convergent. The staudinger ligation was the first example of bioorthogonal ligation reaction.
While the staudinger reaction has first been described a hundred years. Staudinger chemistry, based on the specific crosslinking reaction between azide. Nov 05, 2012 this page was last edited on 23 october 2020, at 17. First phosphine imineforming reaction is conducted involving treatment of the azide with the phosphine. Here, we describe a method for peptide ligation that eliminates the need for a cysteine residue and leaves no residual atoms in the peptide product. He is also known for his discovery of ketenes and of the staudinger reaction. The traceless staudinger ligation is an effective means to synthesize an amide bond. Azobenzene photoswitches for staudingerbertozzi ligation. Jan 12, 2015 hermann staudinger 23 march 1881 8 september 1965 was a german chemist who demonstrated the existence of macromolecules, which he characterized as polymers. Moreover, the staudinger ligation has been used as a synthetic method to construct glycopeptides, microarrays, and functional biopolymers. Bioconjugate techniques is a complete textbook and protocols manual for. Selective traceless staudinger ligation for glycoprotein. That protonation would prevent the desired sn acyl transfer. Here, the coupling of peptides by this reaction was characterized in detail.
Because the staudinger ligation, named by bertozzi after the classic reaction that inspired it, is potentially useful for labeling components of the cell interior such as amino acids from which proteins are assembled previously unobservable interactions in the cell may become accessible. Catalyst free chemoselective dna conjugation by the staudinger ligation and diels alder cycloaddition reaction. Pdf dna conjugation by staudinger ligation anna baccaro. Stereoselective nglycosylation by staudinger ligation. A novel application of the staudinger ligation to access. Cd units in a single step and in aqueousorganic media, under mild conditions and with good yields. Our results provide precedence for the use of this. Watersoluble phosphinothiols for traceless staudinger ligation. Traceless staudinger ligation for bioconjugation of rna. Thus, staudinger ligation could be the method of choice for linking cds.
Synthesis, conformations and guestencapsulation ability malamatenia d. A chemical transformation that permits the selective formation of covalent adducts among richly functionalized biopolymers within a cellular context is presented. Smallmolecule control of protein function through staudinger. Treatment with trifluoroacetic acid tfa results in the simultaneous removal of the borane along with the protecting groups on the peptide side chains, and chemoselective amide.
As a chloride salt, phosphinothiol 17 was an easytohandle, odorfree. This chemoselective reaction allows the ligation of two biomolecules through an azide group and different piii species including phosphines, phosphites or phosphonites. Staudinger azide reduction an overview sciencedirect topics. Jul 25, 2016 using small molecules to control the function of proteins in live cells with complete specificity is highly desirable, but challenging.
Mechanisms and substituent effects of metal free bioorthogonal. Staudinger ligation as a method for bioconjugation van. The staudinger ligation a gift to chemical biology graz. This method has been used along with others to assemble an entire protein. Recent progress using the staudinger ligation for radiolabeling. Chemoselective peptide cyclization by traceless staudinger. D staudinger ligation employing enzymatically synthesized dsdna as shown in fig. Jan 01, 2009 the traceless staudinger ligation has joined the repertoire of ligation methods for the convergent synthesis of proteins. Bjoc staudinger ligation towards cyclodextrin dimers in. The main drawback to this reaction is the acidic or basic conditions required for the hydrolysis of the iminophosphorane intermediate. Staudinger ligation for glycoprotein synthesis creative.
Sloby published staudinger ligation find, read and cite all the research you need on researchgate. Staudinger ligation reaction chemistry thermo fisher scientific id. Thus, the total chemical synthesis of proteins and glycopeptides is enabled overcoming the limitations of native chemical ligation ncl of a cys residue at the. A phosphinobenzenethiol is used to link a thioester and azide in moderate yields. Peptide and protein assembly using the staudinger ligation by l. Bernardes department of chemistry, university of oxford, chemistry research laboratory, 12 mansfield road, ox1 3ta oxford uk, fax. The most prominent chemical ligation techniques click chemistry, native chemical ligation, and staudinger ligation will be discussed. The staudinger ligation is the most efficacious method known for the sitespecific, covalent immobilization of a protein. The staudinger ligation a gift to chemical biology. This work demonstrated the feasibility of photocaging arylphosphines to realize the photo.
Download citation the staudinger ligation the staudinger ligation has been developed as a versatile bioorthogonal conjugation tool for a wide variety of biological applications both in. Mechanisms and substituent effects of metalfree bioorthogonal. A new drugrelease method using the staudinger ligation. Nov 01, 2009 biocompatible, efficient, chemoselective, and catalyst free, the recently introduced staudinger ligation has been used for numerous biological applications, from cell surface modification to protein detection. A powerful ringclosure method to form mediumsized lactams. No other approach enables more rapid immobilization or a higher yield of active protein cf refs 25. Oct 30, 2004 stereoselective methods for the chemical synthesis of. Presenting a wide array of information on chemical ligation one of the more powerful tools for protein and peptide synthesis this book helps readers understand key methodologies and applications that protein therapeutic. Applications in model systems include the modification of azide. In the emerging field of bioorthogonal ligation strategies, the staudinger ligation has set a high standard to which most of the new techniques are often compared.
Since then, the staudinger bertozzi ligation has been used widely in. Staudinger ligation as a method for bioconjugation van berkel. Cell surface engineering by a modified staudinger reaction. This ligation can be visualized using a phosphine that has been modified with the flag epitope tag, a peptide recognized by the. Herein we describe a new way to liberate a drug from a prodrug using the staudinger ligation as the trigger. 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. A new transformation based on the staudinger reaction is described, and its application in the design of a novel switch element to. Protein engineering with the traceless staudinger ligation.
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