Spectrophotometric investigation of the interaction of glutathione with maleimide and N-ethylmaleimide

Friedmann, E.

doi:10.1016/0006-3002(52)90121-2

Cited by 50 publications

(13 citation statements)

References 2 publications

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“…In terms of organic chemistry, the thiol group undergoes nucleophilic substitution with an alkyl halide and the Michael addition to an α,β-unsaturated carbonyl compound, both leading to S-alkylation (Scheme 6). Reagents for the alkylation based on substitution include iodoacetic acid or iodoacetamide (Crestfield et al, 1963), ethyleneimine (Plapp et al, 1967), and those suitable for addition are N-ethylmaleimide (NEM) (Friedmann, 1952), and 4-vinylpyridine (Raftery and Cole, 1966). Interestingly, TCEP used for the reduction of disulfide bond is essentially inert to the addition, hence not reactive with NEM, 4-vinylpyridine, and acrylamides, allowing alkylation of this kind to be carried out without removing TCEP from the reaction mixture.…”

Section: Alkylationmentioning

confidence: 99%

Chemistry in Proteomics: An Interplay between Classical Methods in Chemical Modification of Proteins and Mass Spectrometry at the Cutting Edge

Nakazawa

2006

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This review deals with chemical approaches that facilitate proteomics research. The methods include chemical cleavage of peptide bonds, stepwise degradation of proteins, and site-specific modification enabling the discrimination between functional groups. These reactions could enhance the efficacy of mass spectrometry (MS) for identification, quantification, and sequencing of peptides and proteins. Particular stress is laid on the effect of each modification on mass spectra mainly with respect to the enhancement of peak intensities, the clarity of spectra, and the suppression of ambiguity in peak assignment. Chemical modifications targeting the terminal free amino and carboxyl groups are reviewed, because the charge or isotopic tagging on N-and C-termini has proved effective to identify a protein based on the terminal amino acid sequencing with tandem MS in combination with database searching. Such a tag that is designed to provide a peak with a characteristic sensitivity or pattern could corroborate the implication of mass spectra and facilitate the de novo protein sequencing of mature proteins, which are often subjected to post-translational modifications.

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Section: Alkylationmentioning

confidence: 99%

Chemistry in Proteomics: An Interplay between Classical Methods in Chemical Modification of Proteins and Mass Spectrometry at the Cutting Edge

Nakazawa

2006

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“…The reaction of a free sulfhydryl-group with Nethylmaleimide (NEM) is well known. The formation of the adduct has been followed by spectrophotometric measurement using the decrease of the absorbance at λ = 300 nm [12][13][14]. The cysteine content of several proteins has been determined using this method.…”

Section: Introductionmentioning

confidence: 99%

A simple method for monitoring the cysteine content in synthetic peptides

Horváti

Bősze

Hudecz

et al. 2008

Journal of Peptide Science

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A new method for the determination of the cysteine content of synthetic peptides is described. For this purpose the classical amino acid analysis is not suitable, as cysteine may undergo oxidative dimerisation to cystine during the hydrolysis or evaporation. The intact peptide was reacted with N-ethylmaleimide (NEM) yielding a stable S-(N-ethylsuccinimido)-cysteine derivative, which could be separated by RP-HPLC and characterised by mass spectrometry. For the quantitative determination less than 0.1 microM peptide was sufficient.

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“…30 kDa). [1][2][3] 2) AP OI can also be genetically fused to an enzyme,such as HaloTag, [4] SNAP-tag, [5] CLIP-tag, [6] TMP-tag, [7] or to PYP. [8] These protein tags can then be labelled using highly specific, functionalized irreversible inhibitors.O ther protein tags, such as FAPS [9] and YFAST [10] can be labelled non-covalently with specific ligands.However,the considerable size (ca.…”

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confidence: 99%