Proteomic analysis of liver tissues subjected to early ischemia/reperfusion injury during human orthotopic liver transplantation

Vascotto, Carlo; Cesaratto, Laura; D’Ambrosio, Chiara; Scaloni, Andrea; Avellini, Claudio; Paron, Igor; Baccarani, Umberto; Adani, Gian Luigi; Tiribelli, Claudio; Quadrifoglio, Franco; Tell, Gianluca

doi:10.1002/pmic.200500770

Cited by 54 publications

(58 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Recent reports show that a few proteins are altered in humans undergoing liver transplantation. 25,44,45 In addition, some other proteins were also altered in rabbit hearts following I/R injury. 24,46 However, these studies were conducted using whole cell homogenate without subcellular fractionation and functional characterization of those proteins altered during I/R injury.…”

Section: Discussionmentioning

confidence: 98%

“…24,46 However, these studies were conducted using whole cell homogenate without subcellular fractionation and functional characterization of those proteins altered during I/R injury. 25,44,46 In this study, we specifically aimed at identifying the oxidatively-modified target proteins by focusing on redox modulation of Cys residues and evaluating their functional roles in acute hepatic I/R injury, in the presence or absence of a SOD mimetic/peroxynitrite scavenger. The early time point of reperfusion (2-h) was intentionally chosen to identify early signs of oxidative modifications, which may lead to mitochondrial dysfunction prior to a secondary wave of severe liver damage inflicted by infliltrating immune cells during reperfusion for longer times.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Oxidative Inactivation of Key Mitochondrial Proteins Leads to Dysfunction and Injury in Hepatic Ischemia Reperfusion

et al. 2008

View full text Add to dashboard Cite

SummaryBackground & Aims-Ischemia-reperfusion (I/R) is a major mechanism of liver injury following hepatic surgery or transplantation. Despite numerous reports on the role of oxidative/nitrosative stress and mitochondrial dysfunction in hepatic I/R injury, the proteins that are oxidatively-modified during I/R damage are poorly characterized. This study was aimed at investigating the oxidatively-modified proteins underlying the mechanism for mitochondrial dysfunction in hepatic I/R injury. We also studied the effects of a superoxide dismutase mimetic/peroxynitrite scavenger metalloporphyrin MnTMPyP on oxidatively-modified proteins and their functions.

show abstract

Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

Oxidative Inactivation of Key Mitochondrial Proteins Leads to Dysfunction and Injury in Hepatic Ischemia Reperfusion

et al. 2008

View full text Add to dashboard Cite

show abstract

“…Each sample was analyzed in triplicate, and then a comparative analysis was carried out as previously reported (59). Differential spots from two-dimensional electrophoresis (2-DE) were excised from the gel, triturated, and washed with water.…”

Section: Methodsmentioning

confidence: 99%

“…Differential spots from two-dimensional electrophoresis (2-DE) were excised from the gel, triturated, and washed with water. Proteins were in-gel reduced, S alkylated, and digested with trypsin as previously reported (59). Digest aliquots were removed and subjected to a desalting/concentration step on ZipTip C 18 column (Millipore Corp., Bedford, MA) using acetonitrile as the eluent before matrix-assisted laser desorption ionization-time of flight mass spectrometry analysis.…”

Section: Methodsmentioning

confidence: 99%

APE1/Ref-1 Interacts with NPM1 within Nucleoli and Plays a Role in the rRNA Quality Control Process

Vascotto

Fantini

Romanello

et al. 2009

Molecular and Cellular Biology

Self Cite

220

411

View full text Add to dashboard Cite

APE1/Ref-1 (hereafter, APE1), a DNA repair enzyme and a transcriptional coactivator, is a vital protein in mammals. Its role in controlling cell growth and the molecular mechanisms that fine-tune its different cellular functions are still not known. By an unbiased proteomic approach, we have identified and characterized several novel APE1 partners which, unexpectedly, include a number of proteins involved in ribosome biogenesis and RNA processing. In particular, a novel interaction between nucleophosmin (NPM1) and APE1 was characterized. We observed that the 33 N-terminal residues of APE1 are required for stable interaction with the NPM1 oligomerization domain. As a consequence of the interaction with NPM1 and RNA, APE1 is localized within the nucleolus and this localization depends on cell cycle and active rRNA transcription. NPM1 stimulates APE1 endonuclease activity on abasic double-stranded DNA (dsDNA) but decreases APE1 endonuclease activity on abasic single-stranded RNA (ssRNA) by masking the N-terminal region of APE1 required for stable RNA binding. In APE1-knocked-down cells, pre-rRNA synthesis and rRNA processing were not affected but inability to remove 8-hydroxyguanine-containing rRNA upon oxidative stress, impaired translation, lower intracellular protein content, and decreased cell growth rate were found. Our data demonstrate that APE1 affects cell growth by directly acting on RNA quality control mechanisms, thus affecting gene expression through posttranscriptional mechanisms.APE1/Ref-1 (also called HAP1 or APEX, and here referred to as APE1), the mammalian ortholog of Escherichia coli Xth (exonuclease III), is a vital protein (20) that acts as a master regulator of cellular response to oxidative stress conditions and contributes to the maintenance of genome stability (55, 56). APE1 is involved in both the base excision repair (BER) pathways of DNA lesions, acting as the major apurinic/apyrimidinic (AP) endonuclease, and in transcriptional regulation of gene expression as a redox coactivator of different transcription factors, such as early growth response protein 1 (Egr-1), NF-B, and p53 (55, 56). These two biological activities are located in two functionally distinct protein domains. In fact, the N-terminal region, containing the nuclear localization signal (NLS) sequence, is principally devoted to redox activity through Cys65, while the C-terminal one exerts enzymatic activity on the abasic sites of DNA (56, 63). The protein C terminus is highly conserved during phylogenesis, while the N terminus is not. Except in mammals, which always show a high sequence conservation (more than 90%), and Danio, Drosophila, Xenopus, and Dictyostelium (presenting a sequence identity of less than 40%), the N-terminal region is mostly absent in other organisms. A third APE1 function, which is regulated by Lys6/Lys7 acetylation (7), is indirect binding to the negative calcium response elements (nCaRE) of some promoters (i.e., PTH and APE1 promoters), thus acting as a transcriptional repressor (12,30).In different ...

show abstract

“…Two proteomic studies revealed that the liver proteome changes during OLT; modifications were found to include upregulation of proteins associated with energy metabolism, various metabolic pathways, oxidative-stress antioxidant response, and lipid metabolism) (17,18). Unfortunately, the design of these studies did not allow to answer the question whether or not these modifications would finally result in elevation of oxidative and nitrosative stress.…”

Section: Discussionmentioning

confidence: 99%

Does Oxidative Stress Change During Orthotopic Liver Transplantation?

Tsikas

2017

jcbmr

View full text Add to dashboard Cite

In clinical ischemia/reperfusion injury, damage resulting from oxidative and nitrosative stress is generally considered crucial for graft functioning. Yet, there is reasonable doubt that modern clinical transplantation including orthotopic liver transplantation (OLT) is associated with elevation of oxidative and nitrosative stress upon organ reoxygenation. We measured two biomarkers of oxidative stress, 15(S)-8-iso-prostaglandin F 2α (15(S)-8-iso-PGF 2α ) and cis-epoxyoctadecanoic acid (cis-EpOA), in human plasma during the entire time duration of OLT in nine patients suffering from end-stage liver disease. No considerable concentration changes of 15(S)-8-iso-PGF 2α and cis-EpOA were observed in plasma, indicating lack of oxidative stress. Creatinine-corrected urinary excretion of 15(S)-8-iso-PGF 2α in two urine samples collected 40-60 min and 60-240 min after reperfusion did not increase significantly. Previously, we found in the same patients that nitrosative stress, measured as 3-nitrotyrosine and 3-nitrotyrosinoalbumin, is elevated in the patients but did not change during OLT. 15(S)-8-iso-PGF 2α , cis-EpOA, 3-nitrotyrosine and other widely used biomarkers of oxidative stress, notably malondialdehyde (MDA), are produced both by chemical and enzymatic reactions. This important issue is rarely considered in the area of oxidative and nitrosative stress. In addition, many widely used analytical approaches such as the total antioxidant capacity (TAC) assay to quantitate oxidative and nitrosative stress in health and disease and during clinical transplantation lack reliability. New basic concepts of oxidative and nitrosative stress need to be drafted and implemented in vitro as well as in vivo in experimental and clinical settings.

show abstract

Proteomic analysis of liver tissues subjected to early ischemia/reperfusion injury during human orthotopic liver transplantation

Cited by 54 publications

References 38 publications

Oxidative Inactivation of Key Mitochondrial Proteins Leads to Dysfunction and Injury in Hepatic Ischemia Reperfusion

Oxidative Inactivation of Key Mitochondrial Proteins Leads to Dysfunction and Injury in Hepatic Ischemia Reperfusion

APE1/Ref-1 Interacts with NPM1 within Nucleoli and Plays a Role in the rRNA Quality Control Process

Does Oxidative Stress Change During Orthotopic Liver Transplantation?

Contact Info

Product

Resources

About