Sortase‐Based Bio‐organic Strategies for Macromolecular Synthesis

Haridas, V.; Sadanandan, Sandhya; Dheepthi, N. U.

doi:10.1002/cbic.201402013

Cited by 19 publications

(12 citation statements)

References 78 publications

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“…This complication can be addressed by combining SrtA-mediated ligation with bio-orthogonal cross-linking. To avoid the need for an extra step, target proteins are pre-modified with the bio-orthogonal group using SrtA-mediated site-specific ligation (Haridas et al, 2014). Click chemistry is a versatile bio-orthogonal reaction involving a cycloaddition between an azido group and a terminal alkyne (Beranardin et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

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Streptavidin-hydrogel prepared by sortase A-assisted click chemistry for enzyme immobilization on an electrode

Matsumoto

Isogawa

Tanaka

et al. 2018

Biosensors and Bioelectronics

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A tetrameric streptavidin (SA)-appended LPETG tag was site-specifically linked to azido-containing tri-glycine via sortase A catalysis and the resulting azido-modified SA (SA-N3) was retained in the biotin-binding pocket. SA-N3 was polymerized with dibenzylcyclooctyne-modified branched poly(ethyleneglycol) (DBCO-PEG) using azido-modified branched PEG (N3-PEG) as a spacer via copper-free click chemistry. The resulting SA-based hydrogel exhibited gel-like mechanical properties and could immobilize biotin-modified molecules through biotin-SA affinity. Glucose dehydrogenase (GDH) was immobilized in the SA-based hydrogel, and the hydrogel was then coated on a glassy carbon electrode (GCE) and used for the biocatalytic oxidation of glucose. The designed GCE exhibited better performance and stability compared with GDH chemically adsorbed onto a GCE. In addition, the designed GCE anode and a Pt-carbon cathode were assembled into a glucose/O fuel cell that provided a maximum power density and open circuit voltage of 11.8±0.56µWcm and 0.17V, respectively.

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Section: Introductionmentioning

confidence: 99%

“…Click chemistry is a versatile bio-orthogonal reaction involving a cycloaddition between an azido group and a terminal alkyne (Beranardin et al, 2010). Click handles can be site-specifically incorporated into target molecules using site-specific SrtA-mediated ligation (Witte et al, 2013;Haridas et al, 2014;Wang et al, 2016b).…”

Section: Introductionmentioning

confidence: 99%

Streptavidin-hydrogel prepared by sortase A-assisted click chemistry for enzyme immobilization on an electrode

Matsumoto

Isogawa

Tanaka

et al. 2018

Biosensors and Bioelectronics

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“…Subtiligase is a redesigned peptide ligase based on the modification of the active site of subtilisin. It was engineered by converting catalytic Ser221 to Cys, thereby increasing the ligase activity compared to amidase activity, and Residue Pro225 was converted to Ala to reduce steric assembling (Haridas et al, 2014). Subtiligase facilitates the ligation between a peptide C-terminal ester and a peptide N-terminal α-amine, without requiring a recognition motif at the termini of any reaction partners (Lin and He, 2018) (Figure 3C).…”

Section: Subtiligasementioning

confidence: 99%

From Synthesis to Characterization of Site-Selective PEGylated Proteins

et al. 2019

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Covalent attachment of therapeutic proteins to polyethylene glycol (PEG) is widely used for the improvement of its pharmacokinetic and pharmacological properties, as well as the reduction in reactogenicity and related side effects. This technique named PEGylation has been successfully employed in several approved drugs to treat various diseases, even cancer. Some methods have been developed to obtain PEGylated proteins, both in multiple protein sites or in a selected amino acid residue. This review focuses mainly on traditional and novel examples of chemical and enzymatic methods for site-selective PEGylation, emphasizing in N-terminal PEGylation, that make it possible to obtain products with a high degree of homogeneity and preserve bioactivity. In addition, the main assay methods that can be applied for the characterization of PEGylated molecules in complex biological samples are also summarized in this paper.

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“…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). Furthermore, while this unit does not outline specific procedures for generating each of the conjugates listed here, it should be stressed that most are attainable using simple variations of the Basic and Alternate Protocol outlined above.…”

Section: Commentarymentioning

confidence: 99%

Site‐Specific Protein Labeling via Sortase‐Mediated Transpeptidation

et al. 2017

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Strategies for site-specific protein modification are highly desirable for the construction of conjugates containing non-genetically-encoded functional groups. Ideally, these strategies should proceed under mild conditions, and be compatible with a wide range of protein targets and non-natural moieties. The transpeptidation reaction catalyzed by bacterial sortases is a prominent strategy for protein derivatization that possesses these features. Naturally occurring or engineered variants of sortase A from Staphylococcus aureus catalyze a ligation reaction between a five-amino-acid substrate motif (LPXTG) and oligoglycine nucleophiles. By pairing proteins and synthetic peptides that possess these ligation handles, it is possible to install modifications onto the protein N- or C-terminus in site-specific fashion. As described in this unit, the successful implementation of sortase-mediated labeling involves straightforward solid-phase synthesis and molecular biology techniques, and this method is compatible with proteins in solution or on the surface of live cells.

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Sortase‐Based Bio‐organic Strategies for Macromolecular Synthesis

Cited by 19 publications

References 78 publications

Streptavidin-hydrogel prepared by sortase A-assisted click chemistry for enzyme immobilization on an electrode

Streptavidin-hydrogel prepared by sortase A-assisted click chemistry for enzyme immobilization on an electrode

From Synthesis to Characterization of Site-Selective PEGylated Proteins

Site‐Specific Protein Labeling via Sortase‐Mediated Transpeptidation

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