Backbone cyclization of a recombinant cystine‐knot peptide by engineered Sortase A

Stanger, Karen; Maurer, Till; Kaluarachchi, Harini; Coons, Mary; Franke, Yvonne; Hannoush, Rami N.

doi:10.1016/j.febslet.2014.10.020

Cited by 51 publications

(51 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In a similar fashion Hannoush recently expressed a glutathione S-transferase (GST)-fusion protein containing a linearized version of the cyclotide MCoTI-II (Stanger et al, 2014). To allow the SrtA-mediated backbone cyclization, the SrtA recognition motifs GGG and LPETGG were added the N- and C-termini of the linear cyclotide sequence, respectively.…”

Section: Biological Synthesis Of Cyclotidesmentioning

confidence: 99%

See 1 more Smart Citation

Chemical and Biological Production of Cyclotides

Li¹,

Bi²,

Camarero³

2015

Advances in Botanical Research

View full text Add to dashboard Cite

Cyclotides are fascinating naturally occurring micro-proteins (≈30 residues long) present in several plant families, and display various biological properties such as protease inhibitory, anti-microbial, insecticidal, cytotoxic, anti-HIV and hormone-like activities. Cyclotides share a unique head-to-tail circular knotted topology of three disulfide bridges, with one disulfide penetrating through a macrocycle formed by the two other disulfides and interconnecting peptide backbones, forming what is called a cystine knot topology. This cyclic cystine knot (CCK) framework gives the cyclotides exceptional rigidity, resistance to thermal and chemical denaturation, and enzymatic stability against degradation. Interestingly, cyclotides have been shown to be orally bioavailable, and other cyclotides have been shown to cross the cell membranes. Moreover, recent reports have also shown that engineered cyclotides can be efficiently used to target extracellular and intracellular protein-protein interactions, therefore making cyclotides ideal tools for drug development to selectively target protein-protein interactions. In this work we will review all the available methods for production of these interesting proteins using chemical or biological methods.

show abstract

Section: Biological Synthesis Of Cyclotidesmentioning

confidence: 99%

“…Strategy used for the biosynthesis of a cyclotide using SrtA-induced backbone cyclization (Stanger et al, 2014). …”

Section: Figurementioning

confidence: 99%

Chemical and Biological Production of Cyclotides

Li¹,

Bi²,

Camarero³

2015

Advances in Botanical Research

View full text Add to dashboard Cite

show abstract

“…As the above examples illustrate, recombinant sortase A from S. aureus presents a promising new tool for the (Stanger et al, 2014) backbone cyclization of both proteins and peptides. The enzyme can be produced on a large scale by recombinant expression followed by Ni 2+ -NTA-affinity chromatography.…”

Section: Conclusion: Sortase As a Promising Tool For Backbone Cyclizamentioning

confidence: 99%

“…Preliminary data show that sortase is still active when immobilized to a solid support (Steinhagen et al, 2013), suggesting the feasibility of industrial-scale production of sortase-cyclized proteins and peptides in the near future. Furthermore, a more efficient SrtA variant has recently been produced using a directed evolution strategy (Table 2) (Stanger et al, 2014 SrtA the maximum cyclization yield of 80% was obtained within 48 h, whereas with wild type SrtA only 20% yield was obtained after 96 h.…”

Section: Conclusion: Sortase As a Promising Tool For Backbone Cyclizamentioning

confidence: 99%

Sortase-mediated backbone cyclization of proteins and peptides

Hof

Maňásková²,

Veerman

et al. 2015

Biological Chemistry

View full text Add to dashboard Cite

Backbone cyclization has a profound impact on the biological activity and thermal and proteolytic stability of proteins and peptides. Chemical methods for cyclization are not always feasible, especially for large peptides or proteins. Recombinant Staphylococcus aureus sortase A shows potential as a new tool for the cyclization of both proteins and peptides. In this review, the scope and background of the sortase-mediated cyclization are discussed. High efficiency, versatility, and easy access make sortase A a promising cyclization tool, both for recombinant and chemo-enzymatic production methods.

show abstract

“…[7] Thus,d ue to their favourable properties such as excellent chemo-selectivity,t he use of enzymesf or the head-totail cyclization of peptides has been extensively examined and providesa ne legant link between chemistry and biology. [8] Thec urrently existing set of enzymes used for peptide cyclization is comprisedo fe nzymes such as sortases, [9,10] trypsin, [11] asparaginyl endoproteases (AEP)l ike butelase-1 [12,13] or OaAEP1b [14] and subtilisin variants like peptiligase. [15,16] Most of the enzymatic approachesi nvestigated sufferf rom incomplete ligation reactions and low catalytic efficiency, and in addition leave al igase "footprint", an unavoidable enzyme recognition sequence at the coupling site.I nc ontrast, peptiligase based enzyme variants have recently emerged as av ery powerful tool for traceless (footprint free) enzyme-mediated peptide bond formation.…”

mentioning

confidence: 99%

Omniligase‐1: A Powerful Tool for Peptide Head‐to‐Tail Cyclization

Schmidt

Toplak²,

Quaedflieg³

et al. 2017

Adv Synth Catal

View full text Add to dashboard Cite

Strategies for the efficients ynthesis of peptide macrocyclesh ave been al ong-standing goal. In this paper, we demonstrate the use of the peptide ligase termedo mniligase-1 as av ersatile and broadly applicable enzymatic toolf or peptide cyclization. Severalh ead-to-tail (multi)cyclic peptides have been synthesized, including the cyclotide MCoTI-II. Cyclization ando xidative foldingo ft he cyclotide MCoTI-II were efficiently performed in ao ne-pot reactiono na1 -gram scale.T he native cyclotide was isolated andt he correct disulfide bonding pattern was confirmed by NMRs tructure determination. Furthermore,c ompatibility of chemo-enzymatic peptide synthesis( CEPS) using omniligase1w ith methods such as chemical ligation of peptides onto scaffolds( CLIPS)w as successfully demonstrated by synthesizing ak inase-inhibitor derived tricyclic peptide.O ur studies indicate that the minimal ring size for omniligase-1 mediated cyclization is 11 amino acids,w hereas the cyclization of peptides longer than 12 amino acids proceeds with remarkablee fficiency.I na ddition,s everal macrocycles containing non-peptidic backbones( e.g.,p olyethylene glycol), isopeptide bonds (aminoa cid sidechain attachment) as well as d-amino acids couldbe efficiently cyclized.Keywords: chemo-enzymatic peptide synthesis (CEPS);c yclic peptides; cyclization;c yclotide synthesis;c yclotides;e nzyme catalysis;h ead-to-tail cyclization; ligases;m acrocycles;o mniligase-1;p eptides Peptide macrocyclesr epresent an extremely diverse class of molecules that attracti ncreased attentiona s drug leads and prospective pharmaceuticals,w ith currently over 30 cyclic peptides registered or in clinical trials.[1-3] Togetherw ith linearp eptides, they fill the gap between small-molecule drugs (less than 500 Da) and biologics (over 5000 Da) andh avet he potential to address previously "undruggable" targets. Many cyclicp eptides are characterized by their unique structural and enhancedb iopharmaceutical properties,s uch as an improved metabolic stability due to ar educed sensitivity to proteolytic cleavage.T he increasing number of cyclic peptidesu sed as therapeutics is accompanied by the need for efficient andc osteffectiver outes that enable their synthesis on al arge scale.M oreover, efficients ynthesiso fl arge libraries of cyclic peptides (e.g.,f or screeningp urposes) or of cyclicp eptides containing non-canonical amino acids (e.g., d-o ru nnatural amino acids to increase stability and diversity) is of importance.H owever, efficient cyclization of peptidesu sing traditional synthetic methodsi ss till ac hallenge.[4] Classical coupling reagents are often used to cyclize side-chain protected peptides in anhydrous organic solvents.H owever, heavy dilution to prevent polymerization,r isk of epimerization and ap oors olubility of side-chain protected peptideslimitthis approach, especially for peptides longer than 20 amino acids.[5] Native chemical ligation (NCL) [6] is often used to cyclize unprotected peptides in aqueous solution. However, not all pep...

show abstract

Backbone cyclization of a recombinant cystine‐knot peptide by engineered Sortase A

Cited by 51 publications

References 49 publications

Chemical and Biological Production of Cyclotides

Chemical and Biological Production of Cyclotides

Sortase-mediated backbone cyclization of proteins and peptides

Omniligase‐1: A Powerful Tool for Peptide Head‐to‐Tail Cyclization

Contact Info

Product

Resources

About