Formation of protein cross-links by singlet oxygen-mediated disulfide oxidation
Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
Standard
Formation of protein cross-links by singlet oxygen-mediated disulfide oxidation. / Jiang, Shuwen; Carroll, Luke; Mariotti, Michele; Hagglund, Per; Davies, Michael J.
I: Redox Biology, Bind 41, 101874, 2021.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
Harvard
APA
Vancouver
Author
Bibtex
}
RIS
TY - JOUR
T1 - Formation of protein cross-links by singlet oxygen-mediated disulfide oxidation
AU - Jiang, Shuwen
AU - Carroll, Luke
AU - Mariotti, Michele
AU - Hagglund, Per
AU - Davies, Michael J.
PY - 2021
Y1 - 2021
N2 - Cross-links formed within and between proteins are a major cause of protein dysfunction, and are postulated to drive the accumulation of protein aggregates in some human pathologies. Cross-links can be formed from multiple residues and can be reversible (usually sulfur-sulfur bonds) or irreversible (typically carbon-carbon or carbon-heteroatom bonds). Disulfides formed from oxidation of two Cys residues are widespread, with these formed both deliberately, via enzymatic reactions, or as a result of unintended oxidation reactions. We have recently demonstrated that new protein-glutathione mixed disulfides can be formed through oxidation of a protein disulfide to a thiosulfinate, and subsequent reaction of this species with glutathione. Here we investigate whether similar reactions occur between an oxidized protein disulfide, and a Cys residues on a second protein, to give novel protein cross-links. Singlet oxygen (1O2)-mediated oxidation of multiple proteins (?-lactalbumin, lysozyme, beta-2-microglobulin, C-reactive protein), and subsequent incubation with the Cys-containing protein glyceraldehyde-3-phosphate dehydrogenase (GAPDH), generates inter-protein cross-links as detected by SDSPAGE, immunoblotting and mass spectrometry (MS). The cross-link yield is dependent on the 1O2 concentration, the presence of the original protein disulfide bond, and the free Cys on GAPDH. MS with 18O-labeling has allowed identification of the residues involved in some cases (e.g. Cys25 from the Cys25-Cys80 disulfide in beta2-microglobulin, with Cys149 or Cys244 of GAPDH). The formation of these cross-links results in a loss of GAPDH enzymatic activity. These data provide ?proof-of-concept? for a novel mechanism of protein cross-link formation which may help rationalize the accumulation of cross-linked proteins in multiple human pathologies.
AB - Cross-links formed within and between proteins are a major cause of protein dysfunction, and are postulated to drive the accumulation of protein aggregates in some human pathologies. Cross-links can be formed from multiple residues and can be reversible (usually sulfur-sulfur bonds) or irreversible (typically carbon-carbon or carbon-heteroatom bonds). Disulfides formed from oxidation of two Cys residues are widespread, with these formed both deliberately, via enzymatic reactions, or as a result of unintended oxidation reactions. We have recently demonstrated that new protein-glutathione mixed disulfides can be formed through oxidation of a protein disulfide to a thiosulfinate, and subsequent reaction of this species with glutathione. Here we investigate whether similar reactions occur between an oxidized protein disulfide, and a Cys residues on a second protein, to give novel protein cross-links. Singlet oxygen (1O2)-mediated oxidation of multiple proteins (?-lactalbumin, lysozyme, beta-2-microglobulin, C-reactive protein), and subsequent incubation with the Cys-containing protein glyceraldehyde-3-phosphate dehydrogenase (GAPDH), generates inter-protein cross-links as detected by SDSPAGE, immunoblotting and mass spectrometry (MS). The cross-link yield is dependent on the 1O2 concentration, the presence of the original protein disulfide bond, and the free Cys on GAPDH. MS with 18O-labeling has allowed identification of the residues involved in some cases (e.g. Cys25 from the Cys25-Cys80 disulfide in beta2-microglobulin, with Cys149 or Cys244 of GAPDH). The formation of these cross-links results in a loss of GAPDH enzymatic activity. These data provide ?proof-of-concept? for a novel mechanism of protein cross-link formation which may help rationalize the accumulation of cross-linked proteins in multiple human pathologies.
KW - Disulfide
KW - Singlet oxygen
KW - Protein cross-links
KW - Protein aggregation
KW - Thiol-disulfide exchange
KW - Post-translational modification
KW - Photo-oxidation
KW - INDUCED ELECTROSTATIC STABILIZATION
KW - GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE
KW - MOLECULAR-OXYGEN
KW - SULFENIC ACIDS
KW - MECHANISMS
KW - PHOTOOXIDATION
KW - TYROSINE
KW - BONDS
KW - BETA(2)-MICROGLOBULIN
KW - INACTIVATION
U2 - 10.1016/j.redox.2021.101874
DO - 10.1016/j.redox.2021.101874
M3 - Journal article
C2 - 33601275
VL - 41
JO - Redox Biology
JF - Redox Biology
SN - 2213-2317
M1 - 101874
ER -
ID: 275432311