Sortase-mediated backbone cyclization of proteins and peptides

Abstract

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.

Recent advances in sortase-catalyzed ligation methodology

“…Recent examples include the synthesis of camelid-derived antibody fragment conjugates for the treatment of B-cell lymphoma, the installation of non-isotopically labeled protein domains to facilitate NMR analysis of proteins with limited solubility, the construction of immuno-PET reagents for non-invasive cancer imaging, and the preparation of multifunctional protein nanoparticles [2–5]. This is by no means an exhaustive list, and we refer the reader to other excellent reviews for more comprehensive discussions of sortagging applications [6–10]. Rather than focus on applications, our goal for this review is to provide an overview of advances in sortagging methodology itself.…”

A gold mine for drug discovery: Strategies to develop cyclic peptides into therapies

“…The use of enzymes as biocatalysts in peptide cyclization increasingly attracted attention due to their nontoxic nature, cost‐effectiveness, and high chemoselectivity. With the development of protein engineering biotechnology, a variety of enzymes have been exploited for peptide cyclization, such as butelase, trypsin, protease OaAEP1b, prenyl transferase, PatG, sortase, subtiligase, peptiligase, thioesterase, glutathione S‐transferase, transglutaminase, etc. In 2002, Kohli et al discovered an isolated thioesterase, which displayed the capability of catalyzing the cyclization of linear peptides tethered to a carrier protein via a phosphopantetheine linker.…”

Site‐Specific Protein Labeling via Sortase‐Mediated Transpeptidation

“…If the end goal is selective modification of a specific protein terminus, then such dual-functionalized derivatives should be avoided, as they may be susceptible to intramolecular transpeptidation, giving rise to circular protein products (Antos et al, 2009b; Jia et al, 2014; van ’t Hof, Maňásková, Veerman, & Bolscher, 2015). In certain cases, proteins containing a C-terminal LPXTG site and an N-terminal glycine may be amenable to site-specific modification at both sites.…”

“…Notable examples include the ligation of proteins or peptides to fluorophores (Popp et al, 2007; Yamamoto & Nagamune, 2009), carbohydrates (Samantaray, Marathe, Dasgupta, Nandicoori, & Roy, 2008; Wu, Guo, Wang, Swarts, & Guo, 2010), polymers (Parthasarathy et al, 2007; Qi, Amiram, Gao, McCafferty, & Chilkoti, 2013), solid supports (Chan et al, 2007; Le, Raeeszadeh-Sarmazdeh, Boder, & Frymier, 2015; Sinisi et al, 2012), therapeutics (Beerli, Hell, Merkel, & Grawunder, 2015; Fang et al, 2016), lipids (Antos et al, 2008; Wu et al, 2010), nucleic acids (Koussa, Sotomayor, & Wong, 2014; Pritz et al, 2007), metal chelators (Paterson et al, 2014; Westerlund, Honarvar, Tolmachev, & Eriksson Karlström, 2015), viral particles (Hess et al, 2013; Schoonen et al, 2015), live cells (Popp et al, 2007; Shi et al, 2014; Yamamoto & Nagamune, 2009), or even intramolecular ligations for generating cyclic proteins and peptides (Antos et al, 2009b; Jia et al, 2014; van ’t Hof et al, 2015). A full discussion of all relevant applications is beyond the scope of this unit, and we refer to the reader to reviews on this topic (Haridas, Sadanandan, & Dheepthi, 2014; Popp & Ploegh, 2011; Ritzefeld, 2014; Schmohl & Schwarzer, 2014).…”