Sortase

The efficiency of the sortase-mediated reactions depends on the flexibility and accessibility of the region comprising the recognition motif. Sortase engineering and mutant selection for better motif recognition have been reported [65]. Details regarding sortase mechanism and applications of the enz3mie can be found in an excellent review by Ritzefeld [32]. 

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Kim SH, Shin DS, Oh MN, Chung SC, Lee JS, Oh KB. Inhibition of the bacterial surface protein anchoring transpeptidase sortase by isoquinoline alkaloids. Biosci Biotechnol Biochem 2004 68 421-424. 

Tanaka, T., Yamamoto, T., Tsukiji, S. and Nagamune, T. (2008). Site-specific protein modification on living cells catalyzed by sortase. Chembiochem 9, 802-7. 

Mazmanian, S. K., Ton-That, H., Su, K. and Schneewind, O. (2002). An iron-regulated sortase anchors a class of surface protein during Staphylococcus aureus pathogenesis, Proc. Natl Acad. Sci. USA, 99, 2293-2298. 

The Lipid I and II building blocks may be further elaborated by many other enzymes that modify the sugars or amino acid chains. Branched peptides are added to the Lipid I and II peptide chains either by enzymes that act in an ATP-dependent fashion similar to the MurC-F ligases [39], or by enzymes that add amino acid residues from aminoacyl tRNA intermediates, such as the S. aureus enzymes FemA, FemB and FemX, which form the pentaglycine bridge (O Fig. 3) [36], and the S. pneumoniae enzymes FemM and FemN, which form an L-Ser-L-Ala or L-Ala-L-Ala dipeptide bridge [34,35]. Lipid II is also the substrate for the sortase enzymes that catalyze the attachment of surface proteins for incorporation into peptidoglycan [40]. 

Two new cyclic peptides 299 and 300 were isolated from the culture broth of an actinomycete of the genus Streptomyces isolated from an unidentified marine sponge, their structures were defined to be the dehydroxy and desmethylenyl derivatives of nocardamine, respectively. They exhibited weak inhibition against the enzyme sortase. 

Keywords Click ligation Diels-Alder ligation Expressed protein Ugation GPI anchor MIC ligation Peptide synthesis Protein lipidation Rab Ras Rheb Sortase-mediated protein ligation... 

Sortase- mediated protein ligation Protein with a C-terminal LPXTG motif Native peptide bond with additional amino acid sequence K-Ras4B, lipidated GFP, GPI modified GFP... 

Recently, Guo and co-workers reported sortase-mediated chemoenzymatic synthesis of a GPl-anchored protein [100]. The GPI anchor 84 featuring the common glycan core, a lipid and an additional double glycine unit was coupled to the model protein GFP 83 by SrtA (Scheme 15d). This work has demonstrated that SrtA could accept a complex GPI anchor, suggesting that SrtA-mediated protein ligation is a versatile approach for protein synthesis. 

Scheme 15 Site-specific lipid attachment through sortase-mediated transpeptidation. (a) Mechanism of sortase-mediated ligation, (b) Semisynthesis of lipid modified GFP protein by sortase-mediated ligation, (c) Semisynthesis of lipidated K-Ras4B protein, (d) Semisynthesis of GPI modified GFP protein...

Weiner, E.M., Robson, S., Marohn, M., Clubb, R.T. (2010) The Sortase A enzyme that attaches proteins to the cell wall of Bacillus anthracis contains an unusueil active site architecture. Journal of Biological Chemistry, 285 (30), 23431-23441. 

Genome analyses of various Gram-positive bacteria revealed the presence of pilus gene clusters containing sortases in pathogens such as Clostridium perfringens, enterococci, many streptococcal species, Actinomyces taxon and, recently, also in Lactobacillus rhamnosus GG a human gut commensal and well-studied probiotic strain. ... 

On the other hand, pili were recently also identified in the genome sequences of human gut commensals such as Bifidobacterium and Lactobacillus. In Lactobacillus rhamnosus GG, two sortase-dependent pili, SpaCBA and SpaEF, have been shown to be involved in bacterial attachment to human cells, and bacterial establishment within the human gut... 

Ton-That H, Marraffini LA, Schneewind O. Sortases and pilin elements involved in pilus assembly of Corynebacterium diphtheriae. Md Microbiol. 2004 53(1) 251-261. 

The Gram-positive human pathogen Streptococcus pneumoniae is a frequent causal agent of pneumonia, otitis, and meningitis. Thirty percent of S. pneumoniae clinical isolates are piliated due to carriage of the rlrA operon coding for three sortases, a regulating element and the structural... 

Oh I, Yang W-Y, Chung S-C, Kim T-Y, Oh K-B, Shin J (2011) In vitro sortase A inhibitory and antimicrobial activity of flavonoids isolated from the roots of Sophora flavescens. Arch Pharm Res 34 217... 

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