TY - JOUR
T1 - Protein targets for carbonylation by 4-hydroxy-2-nonenal in rat liver mitochondria
AU - Guo, Jia
AU - Prokai-Tatrai, Katalin
AU - Nguyen, Vien
AU - Rauniyar, Navin
AU - Ughy, Bettina
AU - Prokai, Laszlo
N1 - Funding Information:
This study was supported by the National Institutes of Health , grant number AG025384 . The authors wish to thank to Shastazia White for her excellent technical assistance. Laszlo Prokai is the Robert A Welch Chair in Biochemistry at UNTHSC (endowment number BK-0031).
PY - 2011/10/19
Y1 - 2011/10/19
N2 - Protein carbonylation has been associated with various pathophysiological processes. A representative reactive carbonyl species (RCS), 4-hydroxy-2-nonenal (HNE), has been implicated specifically as a causative factor for the initiation and/or progression of various diseases. To date, however, little is known about the proteins and their modification sites susceptible to "carbonyl stress" by this RCS, especially in the liver. Using chemoprecipitation based on a solid-phase hydrazine chemistry coupled with LC-MS/MS bottom-up approach and database searching, we identified several protein-HNE adducts in isolated rat liver mitochondria upon HNE exposure. The identification of selected major protein targets, such as the ATP synthase β-subunit, was further confirmed by immunoblotting and a gel-based approach in combination with LC-MS/MS. A network was also created based on the identified protein targets, which showed that the main protein interactions were associated with cell death, tumor morphology and drug metabolism, implicating the toxic nature of HNE in the liver mitoproteome. The functional consequence of carbonylation was illustrated by its detrimental impact on the activity of ATP synthase, a representative major mitochondrial protein target for HNE modifications.
AB - Protein carbonylation has been associated with various pathophysiological processes. A representative reactive carbonyl species (RCS), 4-hydroxy-2-nonenal (HNE), has been implicated specifically as a causative factor for the initiation and/or progression of various diseases. To date, however, little is known about the proteins and their modification sites susceptible to "carbonyl stress" by this RCS, especially in the liver. Using chemoprecipitation based on a solid-phase hydrazine chemistry coupled with LC-MS/MS bottom-up approach and database searching, we identified several protein-HNE adducts in isolated rat liver mitochondria upon HNE exposure. The identification of selected major protein targets, such as the ATP synthase β-subunit, was further confirmed by immunoblotting and a gel-based approach in combination with LC-MS/MS. A network was also created based on the identified protein targets, which showed that the main protein interactions were associated with cell death, tumor morphology and drug metabolism, implicating the toxic nature of HNE in the liver mitoproteome. The functional consequence of carbonylation was illustrated by its detrimental impact on the activity of ATP synthase, a representative major mitochondrial protein target for HNE modifications.
KW - 4-Hydroxy-2-nonenal
KW - ATP synthase
KW - Liver
KW - Mitochondria
KW - Protein carbonylation
KW - Redox proteomics
UR - http://www.scopus.com/inward/record.url?scp=80054874658&partnerID=8YFLogxK
U2 - 10.1016/j.jprot.2011.07.009
DO - 10.1016/j.jprot.2011.07.009
M3 - Article
C2 - 21801862
AN - SCOPUS:80054874658
SN - 1874-3919
VL - 74
SP - 2370
EP - 2379
JO - Journal of Proteomics
JF - Journal of Proteomics
IS - 11
ER -