In Situ Protein Labeling in Complex Environments

Tamura, Tomonori; Shigemitsu, Hajime; Hamachi, Itaru

doi:10.1016/b978-0-12-409547-2.12553-3

Cited by 2 publications

(3 citation statements)

References 246 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This situation is changing with the advent of a series of traceless affinity labeling and affinity-guided catalysts (Figure 4a and Figure 5a), both of which our group pioneered. 2,23,57,58,67 These new technologies enable efficient incorporation of various functionalities into endogenous proteins while retaining their original functions in intact cells.…”

Section: ■ Selective Chemical Modification Of Endogenous Proteins In ...mentioning

confidence: 99%

“…In addition, the labeled protein obtained by traditional affinity labeling often loses its native activity because the affinity ligand permanently occupies the ligand-binding pocket, which hampers suitable applications. This situation is changing with the advent of a series of traceless affinity labeling and affinity-guided catalysts (Figure a and Figure a), both of which our group pioneered. ,,,, These new technologies enable efficient incorporation of various functionalities into endogenous proteins while retaining their original functions in intact cells.…”

Section: Selective Chemical Modification Of Endogenous Proteins In Li...mentioning

confidence: 99%

See 1 more Smart Citation

Chemistry for Covalent Modification of Endogenous/Native Proteins: From Test Tubes to Complex Biological Systems

2018

View full text Add to dashboard Cite

Chemical modification of proteins provides powerful tools to realize a broad range of exciting biological applications, including the development of new classes of biopharmaceuticals and functional studies of individual proteins in complex biological systems. Numerous strategies for linking desired chemical probes with target proteins have been developed in the last two decades, with most exploiting genetic protein engineering and/or bio-orthogonal chemistry that utilizes unnatural amino acids incorporated into proteins. Modification of native proteins in test tubes and biological contexts by site-specific and target-selective approaches remains challenging because appropriate organic chemistry to carry out such modifications is currently limited. Nonetheless, a variety of promising strategies have appeared recently that address this grand challenge in chemical biology. These new chemistries yield native protein-based well-defined bioconjugations, specific labeling of endogenous proteins in various biological crude milieus, and the establishment of chemical proteomics as a new research area in protein science. In this Perspective, we focus on recent remarkable progress in chemistry for native protein modification. We survey chemical characteristics of the methods and describe briefly these advanced applications to address unsolved biological issues. Current limitations and future directions of this research field are also discussed.

show abstract

Section: ■ Selective Chemical Modification Of Endogenous Proteins In ...mentioning

confidence: 99%

Section: Selective Chemical Modification Of Endogenous Proteins In Li...mentioning

confidence: 99%

Chemistry for Covalent Modification of Endogenous/Native Proteins: From Test Tubes to Complex Biological Systems

2018

View full text Add to dashboard Cite

show abstract

“…To circumvent these issues, short peptide tag/probe pair systems offer promising alternatives for site-specific protein modification. 5 – 7 In this approach, a peptide tag-fused protein can be labeled with a designed probe through the specific molecular recognition. Since the sizes of the tag and probe are small, the modification is less likely to impair protein function.…”

Section: Introductionmentioning

confidence: 99%

Site-specific covalent labeling of His-tag fused proteins with N-acyl-N-alkyl sulfonamide reagent

Thimaradka

Heroven

et al. 2021

Bioorganic & Medicinal Chemistry

View full text Add to dashboard Cite

The ability to incorporate a desired functionality into proteins of interest in a site-specific manner can provide powerful tools for investigating biological systems and creating therapeutic conjugates. However, there are not any universal methods that can be applied to all proteins, and it is thus important to explore the chemical strategy for protein modification. In this paper, we developed a new reactive peptide tag/probe pair system for site-specific covalent protein labeling. This method relies on the recognition-driven reaction of a peptide tag and a molecular probe, which comprises the lysine-containing short histidine tag (KH6 or H6K) and a binuclear nickel (II)-nitrilotriacetic acid (Ni 2+ -NTA) complex probe containing a lysine-reactive N -acyl- N -alkyl sulfonamide (NASA) group. The selective interaction of the His-tag and Ni 2+ -NTA propeles a rapid nucleophilic reaction between a lysine residue of the tag and the electrophilic NASA group of the probe by the proximity effect, resulting in the tag-site-specific functionalization of proteins. We characterized the reactive profile and site-specificity of this method using model peptides and proteins in vitro , and demonstrated the general utility for production of a nanobody-chemical probe conjugate without compromising its binding ability.

show abstract

In Situ Protein Labeling in Complex Environments

Cited by 2 publications

References 246 publications

Chemistry for Covalent Modification of Endogenous/Native Proteins: From Test Tubes to Complex Biological Systems

Chemistry for Covalent Modification of Endogenous/Native Proteins: From Test Tubes to Complex Biological Systems

Site-specific covalent labeling of His-tag fused proteins with N-acyl-N-alkyl sulfonamide reagent

Contact Info

Product

Resources

About