Reduction of biological substances by water-soluble phosphines: Gamma-globulin (IgG)

Levison, Matthew E.; Josephson, Alan S.; Kirschenbaum, Donald M.

doi:10.1007/bf01899076

Cited by 50 publications

(28 citation statements)

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“…TCEP was first reported by Rauhut et al 25 and was later used in disulfide reduction. 26 In 1991 a modification of the Rauhut preparation of TCEP was reported, 1 along with a mechanistic description of its use in the reduction of certain disulfides (eq 5). 27, 28 Since then, TCEP has been employed in several related synthetic and biological applications.…”

Section: In Situ Reduction Of Cu(ii) For Cu(i)-catalyzed Azidealkyne mentioning

confidence: 99%

“…TCEP was first used in biological disulfide cleavage with human γ-globulin (IgG) in 1969. 26 Since then, TCEP has been widely used for the reduction of disulfide bonds in a variety of peptides and proteins, both in vitro and in vivo. [40][41][42][43][44][45][46] Prior to the introduction of TCEP, disulfide cleavage in biological systems was conducted using thiols such as 2mercaptoethanol (ME), dithioerythritol (DTE), and dithiothreitol (DTT).…”

Section: In Situ Reduction Of Cu(ii) For Cu(i)-catalyzed Azidealkyne mentioning

confidence: 99%

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Tris(2-carboxyethyl)phosphine Hydrochloride

Yost

Knight

Coltart

2008

Encyclopedia of Reagents for Organic Synthesis

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Section: In Situ Reduction Of Cu(ii) For Cu(i)-catalyzed Azidealkyne mentioning

confidence: 99%

Section: In Situ Reduction Of Cu(ii) For Cu(i)-catalyzed Azidealkyne mentioning

confidence: 99%

Tris(2-carboxyethyl)phosphine Hydrochloride

Yost

Knight

Coltart

2008

Encyclopedia of Reagents for Organic Synthesis

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“…Another drawback of DTT, as well as the other dithiols, is their instability in the presence of oxygen; dithiols are easily oxidized in solution, especially in the presence of heavy metals. Phosphines are non-thiol reductants that were first used for the reduction of disulfide bonds in proteins in 1969 when the cleavage of disulfide bonds in gamma globulin by TCEP was reported33. Another phosphine used for the reduction of disulfide bonds in proteins is THP.…”

mentioning

confidence: 99%

Phosphines are ribonucleotide reductase reductants that act via C-terminal cysteines similar to thioredoxins and glutaredoxins

Domkin

Chabes

2014

Sci Rep

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Ribonucleotide reductases (RNRs) catalyze the formation of 2′-deoxyribonucleotides. Each polypeptide of the large subunit of eukaryotic RNRs contains two redox-active cysteine pairs, one in the active site and the other at the C-terminus. In each catalytic cycle, the active-site disulfide is reduced by the C-terminal cysteine pair, which in turn is reduced by thioredoxins or glutaredoxins. Dithiols such as DTT are used in RNR studies instead of the thioredoxin or glutaredoxin systems. DTT can directly reduce the disulfide in the active site and does not require the C-terminal cysteines for RNR activity. Here we demonstrate that the phosphines tris(2-carboxyethyl)phosphine (TCEP) and tris(3-hydroxypropyl)phosphine (THP) are efficient non-thiol RNR reductants, but in contrast to the dithiols DTT, bis(2-mercaptoethyl)sulfone (BMS), and (S)-(1,4-dithiobutyl)-2-amine (DTBA) they act specifically via the C-terminal disulfide in a manner similar to thioredoxin and glutaredoxin. The simultaneous use of phosphines and dithiols results in ~3-fold higher activity compared to what is achieved when either type of reductant is used alone. This surprising effect can be explained by the concerted action of dithiols on the active-site cysteines and phosphines on the C-terminal cysteines. As non-thiol and non-protein reductants, phosphines can be used to differentiate between the redox-active cysteine pairs in RNRs.

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“…The common disulfide-reducing agents (such as dithiothreitol, DTT) are inactivated at acidic pH (91) and, therefore, cannot be used under the slow exchange conditions. The task of reducing disulfides under such conditions can be successfully carried out by another agent, tris(2-carboxyethyl)phosphine (92,93), which has been shown to remain stable and retain its disulfide-reducing capacity at pH as low as 1.5 (91).…”

Section: "Bottom-up" Approaches To Probing the Local Structure Of Intmentioning

confidence: 99%

Mass Spectrometry‐Based Approaches to Study Biomolecular Dynamics: Equilibrium Intermediates

2005

Mass Spectrometry in Biophysics

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In the preceding chapter, we surveyed various MS-based approaches to study higher-order structure of proteins under native conditions. For many decades, such well-defined and highly organized structures were thought of as the most important (if not the only) determinants of protein function. Protein folding was often considered a linear process leading from fully unstructured (and, therefore, dysfunctional) states to the highly organized native (function-competent) state. The advent of NMR has changed our perception of what "functional" protein states are, with the realization that native proteins are very dynamic species. Perhaps the most illustrious examples of the intimate link between protein dynamics and function were found in enzyme catalysis, where the chemical conversion of substrate to product is often driven by relatively small-scale dynamic events within (and often beyond) the active site. It became clear in recent years that large-scale macromolecular dynamics may also be an important determinant of protein function. A growing number of proteins are found to be either partially or fully unstructured under native conditions, and such flexibility (intrinsic disorder) appears to be vital for their function. Proteins that do have native folds under physiological conditions can also exhibit dynamic behavior via local structural fluctuations or by sampling alternative (higher-energy or "activated") conformations transiently. In many cases, such activated (non-native) states are functionally important despite their low Boltzmann weight. Realization of the importance of transient non-native protein structures for their function not only greatly advanced our understanding of processes as diverse as recognition, signaling, and transport Mass

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Reduction of biological substances by water-soluble phosphines: Gamma-globulin (IgG)

Cited by 50 publications

References 0 publications

Tris(2-carboxyethyl)phosphine Hydrochloride

Tris(2-carboxyethyl)phosphine Hydrochloride

Phosphines are ribonucleotide reductase reductants that act via C-terminal cysteines similar to thioredoxins and glutaredoxins

Mass Spectrometry‐Based Approaches to Study Biomolecular Dynamics: Equilibrium Intermediates

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