Selective chemical protein modification

Spicer, Christopher D.; Davis, Benjamin G.

doi:10.1038/ncomms5740

Cited by 867 publications

(790 citation statements)

References 211 publications

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“…[10][11][12][13] It is currently exploited for several applications, ranging from the adaptation of therapeutic biomolecules to improve stability and/or function to the production of novel, functional materials and bottom-up approaches to synthetic biology. 10,14,15 One of the most useful ways that chemical modification is already exploited is for the fluorescent labelling of proteins, which allows direct visualisation of biomolecules in situ, with increasing resolution due to the ongoing development of specialised imaging techniques. 16,17 Indeed, exciting advances in both microscopy and single-molecule detection are currently used for investigating protein distribution, translocation and interactions both in vitro and in vivo.…”

Section: Introductionmentioning

confidence: 99%

How fluorescent labelling alters the solution behaviour of proteins

Quinn

Gnan²,

James

et al. 2015

Phys. Chem. Chem. Phys.

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A complete understanding of the role of molecular anisotropy in directing the self assembly of colloids and proteins remains a challenge for soft matter science and biophysics. For proteins in particular, the complexity of the surface at a molecular level poses a challenge for any theoretical and numerical description. A soft matter approach, based on patchy models, has been useful in describing protein phase behaviour. In this work we examine how chemical modification of the protein surface, by addition of a fluorophore, affects the physical properties of protein solutions. By using a carefully controlled experimental protein model (human gamma-D crystallin) and numerical simulations, we demonstrate that protein solution behaviour defined by anisotropic surface effects can be captured by a custom patchy particle model. In particular, the chemical modification is found to be equivalent to the addition of a large hydrophobic surface patch with a large attractive potential energy well, which produces a significant increase in the temperature at which liquid-liquid phase separation occurs, even for very low fractions of fluorescently labelled proteins. These results are therefore directly relevant to all applications based on the use of fluorescent labelling by chemical modification, which have become increasingly important in the understanding of biological processes and biophysical interactions.

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Section: Introductionmentioning

confidence: 99%

How fluorescent labelling alters the solution behaviour of proteins

Quinn

Gnan²,

James

et al. 2015

Phys. Chem. Chem. Phys.

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“…Although most of the covalent immobilization methods are nonspecific, new strategies have overcome this problem [53] using direct enzyme modification, such as bioeorthogonal reactions for site-specific protein labeling [54,55]. "Click chemistry", for example, has been used extensively used in modern chemistry, especially for immobilizing azido-or alkyne-containing proteins onto alkyne-or azido-coated surfaces, respectively [56].…”

Section: Covalent Linkingmentioning

confidence: 99%

Protein functionalized carbon nanomaterials for biomedical applications

Oliveira

Bisker

Bakh

et al. 2015

Carbon

191

111

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a b s t r a c tSince the discovery of low-dimensional carbon allotropes, there is increasing interest in using carbon nanomaterials for biomedical applications. Carbon nanomaterials have been utilized in the biomedical field for bioimaging, chemical sensing, targeting, delivery, therapeutics, catalysis, and energy harvesting. Each application requires tailored surface functionalization in order to take advantage of a desired property of the nanoparticles. Herein, we review the surface immobilization of bio-molecules, including proteins, peptides, and enzymes, and present the recent advances in synthesis and applications of these conjugates. The carbon scaffold and the biological moiety form a complex interface which presents a challenge for achieving efficient and robust binding while preserving biological activity. Moreover, some applications require the utilization of the protein-nanocarbon system in a complex environment that may hinder its performance or activity. We analyze different strategies to overcome these challenges when using carbon nanomaterials as protein carriers, explore various immobilization techniques along with characterization methods, and present recent demonstrations of employing these systems for biomedical applications. Finally, we consider the challenges and future directions of this field.

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“…Both compounds decompose, however, into the bioactive methylene quinuclidinone (MQ), which contains a Michael acceptor group that reacts with nucleophilic thiols through a 1,4-addition (12). The nucleophilic -SH of cysteine can be selectively modified by various electrophiles, such as maleimides, iodoacetamides, α-halocarbonyls, Michael acceptors, activated thiols, and methane/ phenylthiosulfonates (13). For a number of other thiol reactive compounds, such as the maleimide MIRA-3, or STIMA-1, similar biological effects were observed (11,14).…”

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confidence: 99%

2-Sulfonylpyrimidines: Mild alkylating agents with anticancer activity toward p53-compromised cells

Bauer

Joerger

Fersht

2016

Proc. Natl. Acad. Sci. U.S.A.

103

113

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The tumor suppressor p53 has the most frequently mutated gene in human cancers. Many of p53’s oncogenic mutants are just destabilized and rapidly aggregate, and are targets for stabilization by drugs. We found certain 2-sulfonylpyrimidines, including one named PK11007, to be mild thiol alkylators with anticancer activity in several cell lines, especially those with mutationally compromised p53. PK11007 acted by two routes: p53 dependent and p53 independent. PK11007 stabilized p53 in vitro via selective alkylation of two surface-exposed cysteines without compromising its DNA binding activity. Unstable p53 was reactivated by PK11007 in some cancer cell lines, leading to up-regulation of p53 target genes such as p21 and PUMA. More generally, there was cell death that was independent of p53 but dependent on glutathione depletion and associated with highly elevated levels of reactive oxygen species and induction of endoplasmic reticulum (ER) stress, as also found for the anticancer agent PRIMA-1MET(APR-246). PK11007 may be a lead for anticancer drugs that target cells with nonfunctional p53 or impaired reactive oxygen species (ROS) detoxification in a wide variety of mutant p53 cells.

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Selective chemical protein modification

Cited by 867 publications

References 211 publications

How fluorescent labelling alters the solution behaviour of proteins

How fluorescent labelling alters the solution behaviour of proteins

Protein functionalized carbon nanomaterials for biomedical applications

2-Sulfonylpyrimidines: Mild alkylating agents with anticancer activity toward p53-compromised cells

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