LC/MS Analysis of Proteins and Peptides in Drug Discovery

Chen, Guodong; Liu, Yan‐Hui; Pramanik, Birendra N.

doi:10.1002/9780470087954.ch19

Cited by 3 publications

(3 citation statements)

References 115 publications

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“…Since the elemental difference between F1108 and F1064 is ϩCϩ2O, and between F1108 and F1036 is ϩ2Cϩ3O, the ions at m/z 1065.2, 1047.2, and 1037.2 are consistent with losses of CO 2 , CO 2 ϩ H 2 O, and CO 2 ϩ CO, respectively. The loss of CO 2 has been shown to be a typical feature in the product ion spectra of pyruvic acid derived compounds [31,32]. Based on this and the limited number of structural possibilities for addition of two carbon and three oxygen atoms to F1036, we propose that F1108 has a glyoxylic N-terminus: HO-CO-COLeu-, as shown in Scheme 1.…”

Section: Reaction Products From the Carboxy Side Of P16mentioning

confidence: 95%

A tris (2-carboxyethyl) phosphine (TCEP) related cleavage on cysteine-containing proteins

Liu

O’Mara

Warrack

et al. 2010

J. Am. Soc. Mass Spectrom.

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Introduced in the late 1980s as a reducing reagent, Tris (2-carboxyethyl) phosphine (TCEP) has now become one of the most widely used protein reductants. To date, only a few studies on its side reactions have been published. We report the observation of a side reaction that cleaves protein backbones under mild conditions by fracturing the cysteine residues, thus generating heterogeneous peptides containing different moieties from the fractured cysteine. The peptide products were analyzed by high performance liquid chromatography and tandem mass spectrometry (LC/MS/MS). Peptides with a primary amine and a carboxylic acid as termini were observed, and others were found to contain amidated or formamidated carboxy termini, or formylated or glyoxylic amino termini. Formamidation of the carboxy terminus and the formation of glyoxylic amino terminus were unexpected reactions since both involve breaking of carbon-carbon bonds in cysteine.

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Section: Reaction Products From the Carboxy Side Of P16mentioning

confidence: 95%

A tris (2-carboxyethyl) phosphine (TCEP) related cleavage on cysteine-containing proteins

Liu

O’Mara

Warrack

et al. 2010

J. Am. Soc. Mass Spectrom.

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“…However, the loss of all membrane proteins, the appearance of multiple proteins in a single spot, and the presence of a single protein in multiple spots constitute some important limitations in the use of this technique. As a result, the most frequently used method for measuring cellular proteins has become liquid chromatography coupled with mass spectrometry (LC-MS or LC-MS/MS), because of its sensitivity, selectivity, accuracy, speed, and throughput [83][84][85][86]. Hence, proteomics has recently been applied to study the metabolic pathways' modifications, biodistribution, and bioaccumulation caused by plastic molecules in aquatic environments, showing huge potential [87].…”

Section: Proteomicsmentioning

confidence: 99%

Polystyrene Micro- and Nanoplastics (PS MNPs): A Review of Recent Advances in the Use of -Omics in PS MNP Toxicity Studies on Aquatic Organisms

Eliso,

Billè,

Cappello

et al. 2024

Fishes

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In recent years, micro- and nanoplastics (MNPs) have represented an emerging threat for the aquatic environment due to their persistence and widespread distribution. Indeed, their small size and increased surface area lead to a high biological reactivity, which can be crucial for the health status of biota. To date, several studies have investigated lethal and sublethal endpoints of MNPs, while one of the main challenges is to understand their mechanisms of toxicity in a comprehensive manner. With this aim, the field of aquatic ecotoxicology can now benefit from the -omics approaches. This review focuses on the recent advances related to the use of transcriptomics, proteomics, and metabolomics to deeply investigate the molecular and biochemical responses of aquatic organisms, both fishes and aquatic invertebrates, to pristine polystyrene (PS) MNPs. The literature reviewed revealed that transcriptomics and metabolomics are the most frequently used -omic approaches. Overall, the studies taken into consideration shed light on the events triggered by PS MNPs at molecular and cellular levels, identifying as mechanisms of toxicity the pathways involved in oxidative stress, energy metabolism, immune response, and the nervous system. Future studies should therefore focus on the use and integration of multi-omics approaches for a more comprehensive understanding of the mechanisms involved in MNPs toxicity.

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“…Additionally, LC–MS offers a great potential for the analysis of proteins that are both intact and proteolyzed. The latest approach represents the clear majority of LC–MS applications that are commonly performed for pharmacokinetic and toxicokinetic studies. − …”

Section: Introductionmentioning

confidence: 99%

Characterization and Detection of Erythropoietin Fc Fusion Proteins Using Liquid Chromatography–Mass Spectrometry

Mesonzhnik

Postnikov²,

Appolonova

et al. 2017

J. Proteome Res.

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Erythropoietin Fc (EPO-Fc) fusion proteins are potential drug candidates that have been designed for the treatment of anemia in humans by stimulating erythrocyte production. Such compounds can be considered performance-enhancing agents that may be used by athletes in endurance sports. This study describes the primary structure of commercially available EPO-Fc based on comprehensive liquid chromatography coupled with mass spectrometry (LC-MS) analysis. A bottom-up approach and the intact molecular weight (MW) measurement of deglycosylated protein and its IdeS proteolytic fractions was used to determine the amino acid sequence of EPO-Fc. Using multiple proteases, peptides covering unknown fusion breakpoints (spacer peptides) were identified. We demonstrated that "spacer peptides" can be used in the determination of EPO-Fc fusion proteins in biological samples using common LC-tandem MS methods.

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LC/MS Analysis of Proteins and Peptides in Drug Discovery

Cited by 3 publications

References 115 publications

A tris (2-carboxyethyl) phosphine (TCEP) related cleavage on cysteine-containing proteins

A tris (2-carboxyethyl) phosphine (TCEP) related cleavage on cysteine-containing proteins

Polystyrene Micro- and Nanoplastics (PS MNPs): A Review of Recent Advances in the Use of -Omics in PS MNP Toxicity Studies on Aquatic Organisms

Characterization and Detection of Erythropoietin Fc Fusion Proteins Using Liquid Chromatography–Mass Spectrometry

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