Study of primary amines for nucleophilic cleavage of cyanylated cystinyl proteins in disulfide mass mapping methodology

Gallegos-Pérez, José-Luis; Rangel-Ordóñez, Laura; Bowman, Stephen Robert; Ngowe, Charles O.; Watson, J. Throck

doi:10.1016/j.ab.2005.08.003

Cited by 12 publications

(6 citation statements)

References 27 publications

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“…Identifying cleavage products by mass spectrometry may be facilitated by cleavage in both ammonia and in the presence of methylamine in parallel reactions, as products with a defined mass difference, i.e. 14 Da, are formed under these conditions (51).…”

Section: Discussionmentioning

confidence: 99%

Disulfide Bond Structure and Domain Organization of Yeast β(1,3)-Glucanosyltransferases Involved in Cell Wall Biogenesis

Popolo

Ragni

Carotti

et al. 2008

Journal of Biological Chemistry

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The Gel/Gas/Phr family of fungal ␤(1,3)-glucanosyltransferases plays an important role in cell wall biogenesis by processing the main component ␤(1,3)-glucan. Two subfamilies are distinguished depending on the presence or absence of a C-terminal cysteine-rich domain, denoted "Cys-box." The N-terminal domain (NtD) contains the catalytic residues for transglycosidase activity and is separated from the Cys-box by a linker region. To obtain a better understanding of the structure and function of the Cys-box-containing subfamily, we identified the disulfide bonds in Gas2p from Saccharomyces cerevisiae by an improved mass spectrometric methodology. We mapped two separate intra-domain clusters of three and four disulfide bridges. One of the bonds in the first cluster connects a central Cys residue of the NtD with a single conserved Cys residue in the linker. Site-directed mutagenesis of the Cys residue in the linker resulted in an endoplasmic reticulum precursor that was not matured and underwent a gradual degradation. The relevant disulfide bond has a crucial role in folding as it may stabilize the NtD and facilitate its interaction with the C-terminal portion of a Gas protein. The four disulfide bonds in the Cys-box are arranged in a manner consistent with a partial structural resemblance with the plant X8 domain, an independent carbohydratebinding module that possesses only three disulfide bonds. Deletion of the Cys-box in Gas2 or Gas1 proteins led to the formation of an NtD devoid of any enzymatic activity. The results suggest that the Cys-box is required for proper folding of the NtD and/or substrate binding.In yeast and fungal cells, the cell wall determines cell shape, protects cells from lysis, and constitutes a permeability barrier to the uptake of exogenous substances. In fungal pathogens, the cell wall is also the interface for interactions with the host and is required for cell adhesion and virulence. In yeast, the extracellular matrix is formed and strengthened by the correct crosslinking of several polymers, mannoproteins, glucans, and chitin (1, 2) where ␤(1,3)-glucan is the most abundant polymer forming a helical, branched structure that is probably responsible for the elastic properties of the cell wall (3, 4). A plasma membrane ␤(1,3)-glucan synthase complex extrudes the polymer outside the cell, and other families of enzymes are responsible for its incorporation into the cell wall and for the anchoring of the other components in a way that is still poorly understood (1). The Gel/Gas/Phr family of proteins plays an essential role in cell wall biogenesis acting as ␤(1,3)-glucan processing enzymes (5, 6). In vitro these proteins catalyze a ␤(1,3)-glucanosyltransferase reaction that consists of the cleavage of an internal glycosidic linkage of a ␤(1,3)-glucan chain, the release of the reducing portion, and the transfer of the new reducing end to the nonreducing end of another acceptor ␤(1,3)-glucan (5, 6). Thus ␤(1,3)-glucanosyltransferases act similarly to glycoside hydrolases, but a carbohydrate functi...

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Section: Discussionmentioning

confidence: 99%

Disulfide Bond Structure and Domain Organization of Yeast β(1,3)-Glucanosyltransferases Involved in Cell Wall Biogenesis

Popolo

Ragni

Carotti

et al. 2008

Journal of Biological Chemistry

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“…The assignment of disulfide bonds can contribute to the understanding of biological function of proteins. However, unambiguous assignment of disulfide bonds is far from trivial and still presents a challenge to analytical chemists [1][2][3]. The most common complication encountered is solution-phase disulfide bond scrambling that changes the original disulfide bond pattern and results in erroneous data.…”

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confidence: 99%

“…A free thiol-containing antibiotic metabolite, desfuroylceftiofur (DFC), was used to derivatize peptides into disulfide-linked bioactive compounds. The MS/MS analysis of one derivative, [vasopressin(SS-DFC) 2 ϩ H] ϩ by CAD resulted in loss of one and two DFC moieties through either homolytic or heterolytic dissociation of the disulfide bonds, including hydrogen migration in the latter case. In contrast, the homolytic cleavage resulted in the formation of an odd-electron product ion and a neutral fragment.…”

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confidence: 99%

Gas-phase scrambling of disulfide bonds during matrix-assisted laser desorption/ionization mass spectrometry analysis

Zhao¹,

Almaraz

Fan

et al. 2009

J. Am. Soc. Mass Spectrom.

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Evidence for photo-induced radical disulfide bond scrambling in the gas phase during matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is described. The phenomenon was observed during the analysis of tryptic peptides from insulin and was confirmed in the determination of disulfide bonds in the rhamnose-binding lectin SEL24K from the Chinook salmon Oncorhynchus tshawytscha. A possible mechanism for this surprising scrambling is proposed. Despite this finding, the disulfide bond pattern in SEL24K was assigned unambiguously by a multi-enzyme digestion strategy in combination with MALDI mass spectrometry. The pattern was found to be symmetrical in the tandem repeat sequence of SEL24K. To the best of our knowledge, this is the first report of disulfide bond scrambling in the gas phase during MALDI-MS analysis. This observation has important ramifications for unambiguous assignment of disulfide bonds. (J Am Soc

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“…Analyses of the undesired materials in both cases indicated that one potential reason for the unsatisfactory outcome was hydrazinolysis‐mediated loss of the cyano group on the cysteine residue, followed by disulfide bond formation between the S ‐sulfonylcysteine and regenerated cysteine residues . Instead of the S ‐sulfonate group, we initially examined disulfide‐type protections such as S–S ‐methyl and S–S ‐ tert ‐butyl groups, because methylamine‐mediated aminolysis of a disulfide‐bond‐containing S ‐cyanopeptide was reported to be possible . However, the use of disulfide protection for hydrazinolysis failed to afford the desired peptides preferentially.…”

Section: Resultsmentioning

confidence: 99%

Preparation of peptide thioesters from naturally occurring sequences using reaction sequence consisting of regioselective S‐cyanylation and hydrazinolysis

et al. 2016

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The vital roles of peptide/protein thioesters in protein chemistry, including chemical or semi-synthesis of proteins, have encouraged studies on the development of methods for the preparation of such chemical units. Biochemical protocols using intein or sortase have proved to be useful in protein chemistry as methods suitable for naturally occurring sequences, including recombinant proteins. Although chemical protocols are potential options for thioester preparation, only a few are applicable to naturally occurring sequences, because standard chemical protocols require an artificial chemical device for producing thioesters. In this context, the chemical preparation of thioesters based on a reaction sequence consisting of regioselective S-cyanylation and hydrazinolysis was investigated. Regioselective S-cyanylation, which is required for cysteine-containing thioesters, was achieved with the aid of a zinc-complex formation of a CCHH-type zinc-finger sequence. Free cysteine residues that are not involved in complex formation were selectively protected with a 6-nitroveratryl group followed by S-cyanylation of the zinc-binding cysteine. Hydrazinolysis of the resulting S-cyanopeptide and subsequent photo-removal of the 6-nitroveratryl group yielded the desired peptide hydrazide, which was then converted to the corresponding thioester. The generated thioester was successfully used in N-to-C-directed one-pot/sequential native chemical ligation using an N-sulfanylethylanilide peptide to give a 64-residue peptide toxin. © 2015 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 106: 531-546, 2016.

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Study of primary amines for nucleophilic cleavage of cyanylated cystinyl proteins in disulfide mass mapping methodology

Cited by 12 publications

References 27 publications

Disulfide Bond Structure and Domain Organization of Yeast β(1,3)-Glucanosyltransferases Involved in Cell Wall Biogenesis

Disulfide Bond Structure and Domain Organization of Yeast β(1,3)-Glucanosyltransferases Involved in Cell Wall Biogenesis

Gas-phase scrambling of disulfide bonds during matrix-assisted laser desorption/ionization mass spectrometry analysis

Preparation of peptide thioesters from naturally occurring sequences using reaction sequence consisting of regioselective S‐cyanylation and hydrazinolysis

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