Abstract
The formation of disulfide bonds is critical for stabilizing protein structures and maintaining protein functions. It is important to understand the linkages between multiple cysteine residues within a protein. In this review, the analytical approaches using mass spectrometry (MS) for disulfide linkage assignment are classified and discussed. Enzymatic digestion under appropriate conditions followed by various MS detection strategies remains the primary method for cysteine linkage analysis. In-source decay (ISD) and electron transfer dissociation (ETD) have been used to generate significant peptide signals that indicate the identities of peptides involved in disulfide bonds. In addition, chemical labeling and software algorithms were also developed to facilitate the automation of disulfide bond analysis. For proteins with complex disulfide structure, methods involving partial reduction coupled with differential alkylation were demonstrated to be useful. In the past two decades, MS has become one of the most valuable tools for protein disulfide bond analysis. It provides irreplaceable information including the peptide backbone sequences as well as the cysteine connection pattern when coupling with appropriate sample preparations. The related approaches with their unique features can be applied for different aims such as structural characterization or functional studies of proteins.
| Original language | English |
|---|---|
| Pages (from-to) | 257-268 |
| Number of pages | 12 |
| Journal | Reviews in Analytical Chemistry |
| Volume | 32 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - 2013 Nov 1 |
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Keywords
- Disulfide bond
- Mass spectrometry
- Protein pharmaceuticals
ASJC Scopus subject areas
- Analytical Chemistry
Cite this
Mass spectrometry-based strategies for protein disulfide bond identification. / Tsai, Pei Lun; Chen, Sung-Fang; Huang, Sheng Yu.
In: Reviews in Analytical Chemistry, Vol. 32, No. 4, 01.11.2013, p. 257-268.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Mass spectrometry-based strategies for protein disulfide bond identification
AU - Tsai, Pei Lun
AU - Chen, Sung-Fang
AU - Huang, Sheng Yu
PY - 2013/11/1
Y1 - 2013/11/1
N2 - The formation of disulfide bonds is critical for stabilizing protein structures and maintaining protein functions. It is important to understand the linkages between multiple cysteine residues within a protein. In this review, the analytical approaches using mass spectrometry (MS) for disulfide linkage assignment are classified and discussed. Enzymatic digestion under appropriate conditions followed by various MS detection strategies remains the primary method for cysteine linkage analysis. In-source decay (ISD) and electron transfer dissociation (ETD) have been used to generate significant peptide signals that indicate the identities of peptides involved in disulfide bonds. In addition, chemical labeling and software algorithms were also developed to facilitate the automation of disulfide bond analysis. For proteins with complex disulfide structure, methods involving partial reduction coupled with differential alkylation were demonstrated to be useful. In the past two decades, MS has become one of the most valuable tools for protein disulfide bond analysis. It provides irreplaceable information including the peptide backbone sequences as well as the cysteine connection pattern when coupling with appropriate sample preparations. The related approaches with their unique features can be applied for different aims such as structural characterization or functional studies of proteins.
AB - The formation of disulfide bonds is critical for stabilizing protein structures and maintaining protein functions. It is important to understand the linkages between multiple cysteine residues within a protein. In this review, the analytical approaches using mass spectrometry (MS) for disulfide linkage assignment are classified and discussed. Enzymatic digestion under appropriate conditions followed by various MS detection strategies remains the primary method for cysteine linkage analysis. In-source decay (ISD) and electron transfer dissociation (ETD) have been used to generate significant peptide signals that indicate the identities of peptides involved in disulfide bonds. In addition, chemical labeling and software algorithms were also developed to facilitate the automation of disulfide bond analysis. For proteins with complex disulfide structure, methods involving partial reduction coupled with differential alkylation were demonstrated to be useful. In the past two decades, MS has become one of the most valuable tools for protein disulfide bond analysis. It provides irreplaceable information including the peptide backbone sequences as well as the cysteine connection pattern when coupling with appropriate sample preparations. The related approaches with their unique features can be applied for different aims such as structural characterization or functional studies of proteins.
KW - Disulfide bond
KW - Mass spectrometry
KW - Protein pharmaceuticals
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U2 - 10.1515/revac-2013-0011
DO - 10.1515/revac-2013-0011
M3 - Article
AN - SCOPUS:84888273200
VL - 32
SP - 257
EP - 268
JO - Reviews in Analytical Chemistry
JF - Reviews in Analytical Chemistry
SN - 0793-0135
IS - 4
ER -