共查询到20条相似文献,搜索用时 62 毫秒
1.
Wan C La Y Zhu H Yang Y Jiang L Chen Y Feng G Li H Sang H Hao X Zhang G He L 《Amino acids》2007,32(1):101-108
Summary. In this study we focused on detecting schizophrenia related changes of plasma proteins using proteomic technology and examining
the relation between schizophrenia and haptoglobin (Hp) genotype. We investigated plasma proteins from schizophrenic subjects (n = 42) and healthy controls (n = 46) by two-dimensional gel electrophoresis (2-DE) in combination with mass spectrometry. To further reveal the genetic
relationship between acute phase proteins (APPs) and schizophrenia disease, we tested Hp α1/Hp α2 (Hp 1/2) polymorphism and two single nucleotide polymorphisms (SNPs) of Hp, rs2070937 and rs5473, for associations with schizophrenia in the Chinese Han population. With the relatively high number
of samples for 2-DE work, we found that four proteins in the family of positive APPs were all up-regulated in patients. In
genetic association study, we found significant associations existing between schizophrenia and Hp polymorphisms, Hp 1/2 and rs2070937 variants. Schizophrenia is accompanied by both an altered expression of Hp protein and a different genotype
distribution of Hp gene, demonstrating that Hp is associated with schizophrenia. The results from proteomic and genomic aspects both indicate
that acute phase reaction is likely to be an aetiological agent in the pathophysiology of schizophrenia, but not just an accompanying
symptom. The positive APPs are schizophrenic related proteins, with the highly concordant results on four positive APPs.
The first two authors contributed equally. 相似文献
2.
Identification of tumor-associated plasma biomarkers using proteomic techniques: from mouse to human 总被引:6,自引:0,他引:6
Juan HF Chen JH Hsu WT Huang SC Chen ST Yi-Chung Lin J Chang YW Chiang CY Wen LL Chan DC Liu YC Chen YJ 《Proteomics》2004,4(9):2766-2775
In an effort to identify tumor-associated proteins from plasma of tumor-bearing mice that may be used as diagnostic biomarkers, we developed a strategy that combines a tumor xenotransplantation model in nude mice with comparative proteomic technology. Five human cancer cell lines (SC-M1, HONE-1, CC-M1, OECM1, GBM 8401) derived from stomach, nasopharyngeal, colon, oral and brain cancers were subcutaneously inoculated into nude mice and compared to control nude mice injected with phosphate-buffered saline. One month later, plasma from mice inoculated with cancer cells was collected for proteomic analysis using two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS). Comparison of plasma 2-DE maps from tumor-bearing mice with those produced from control mice revealed the overexpression of several mouse acute phase proteins (APPs) such as haptoglobin. Another APP, serum amyloid A (SAA), was found only in mice bearing tumors induced by the stomach cancer cell line SC-M1, which has not previously been demonstrated in xenotransplatation experiment. Furthermore, by using immunohistochemistry, SAA and haptoglobin were found to originate from the mouse hosts and not from the human cancer cell line donors. The protein alterations were further confirmed on patients with stomach cancers where up-regulated levels of SAA were also observed. These results indicate that APPs may be used as nonspecific tumor-associated serum markers. SAA in particular may serve as a potential marker for detecting stomach cancer. Taken together, the combination of the xenotransplatation model in nude mice and proteomics analysis provided a valuable impact for clinical applications in cancer diagnostics. In addition, our findings demonstrate that a panel of APPs might serve as screening biomarkers for early cancer detection. 相似文献
3.
Young Yil Bahk Byoung-Kuk Na Shin-Hyeong Cho Jung-Yeon Kim Kook-Jin Lim Tong-Soo Kim 《The Korean journal of parasitology》2010,48(3):203-211
Advancements in the field of proteomics have provided great opportunities for the development of diagnostic and therapeutic tools against human diseases. In this study, we analyzed haptoglobin and amyloid A protein levels of vivax malaria patients with combinations of depletion of the abundant plasma proteins, 2-dimensional gel electrophoresis (2-DE), image analysis, and mass spectrometry in the plasma between normal healthy donors and vivax malaria patients. The results showed that the expression level of haptoglobin had become significantly lower or undetectable in the plasma of vivax malaria patients due to proteolytic cleavage when compared to healthy donors on 2-DE gels. Meanwhile, serum amyloid A protein was significantly increased in vivax malaria patient''s plasma with high statistical values. These 2 proteins are common acute phase reactants and further large scale evaluation with a larger number of patient''s will be necessary to establish the possible clinical meaning of the existential changes of these proteins in vivax malaria patients. However, our proteomic analysis suggests the feasible values of some plasma proteins, such as haptoglobin and serum amyloid A, as associating factor candidates for vivax malaria. 相似文献
4.
Attenuation of the growth hormone (GH)/insulin-like growth factor-1 (IGF-1) axis results in extended lifespan in many organisms
including mice. Conversely, GH transgenic mice have excess GH action and die prematurely. We have studied bovine (b) GH transgenic
mice (n = 9) and their wild type (WT) littermates (n = 8) longitudinally and have determined several age-related changes. Compared to WT mice, bGH mice lost fat mass, became
hypoglycemic and had lower insulin levels at older ages despite being hyperinsulinemic when young. To examine plasma protein
differences in bGH mice relative to controls, samples at 2, 4, 8, 12 and 16 months of age were analyzed by two-dimensional
gel electrophoresis followed by identification using mass spectrometry. We found several differences in plasma proteins of
bGH mice compared to controls, including increased apolipoprotein E (five isoforms), haptoglobin (four isoforms) and mannose-binding
protein-C (one out of three isoforms), and decreased transthyretin (six isoforms). In addition, clusterin (two out of six
isoforms) and haptoglobin (four isoforms) were up-regulated in bGH mice as a function of age. Finally, alpha-2 macroglobulin
(seven isoforms) was altered in an isoform-specific manner with two isoforms increased and two decreased in bGH mouse plasma
compared to controls. In conclusion, identification of these proteins suggests that bGH mice exhibit an increased inflammatory
state with an adverse lipid profile, possibly contributing to their diminished life expectancy. Also, these newly discovered
plasma proteins may be indicative or ‘biomarkers’ of a shortened lifespan. 相似文献
5.
In SJL mice, growth of RcsX lymphoma cells induces an inflammatory response by stimulating V(beta)16+ T cells. During inflammation, various serum protein levels can increase (e.g., acute phase reactants) or decrease (e.g., albumin), and most of these altered proteins are thus potential biomarkers. Although blood plasma is a valuable and promising sample for biomarker discovery for diseases or for novel drug targets, its proteome is complex. To address this, we have focused on a comprehensive comparison of the plasma proteomes from normal and RcsX-tumor-bearing SJL mice using the 1D-Gel-LC-MS/MS method after removing albumin and immunoglobulins. This analysis resulted in the identification of a total of 1079 nonredundant mouse plasma proteins; more than 480 in normal and 790 in RcsX-tumor-bearing SJL mouse plasma. Of these, only 191 proteins were found in common. The molecular weights ranged from 2 to 876 kDa, covering the pI values between 4.22 and 12.09, and included proteins with predicted transmembrane domains. By comparing the plasma proteomic profile of normal and RcsX-tumor-bearing SJL mice, we found significant changes in the levels of many proteins in RcsX-tumor-bearing mouse plasma. Most of the up-regulated proteins were identified as acute-phase proteins (APPs). Also, several unique proteins i.e., haptoglobin, proteosome subunits, fetuin-B, 14-3-3 zeta, MAGE-B4 antigen, etc, were found only in the tumor-bearing mouse plasma; either secreted, shed by membrane vesicles, or externalized due to cell death. These results affirm the effectiveness of this approach for protein identification from small samples, and for comparative proteomics in potential animal models of human disorders. 相似文献
6.
Gamberi T Puglia M Guidi F Magherini F Bini L Marzocchini R Modesti A Modesti PA 《Molecular bioSystems》2011,7(10):2855-2862
Our aim was to identify the key proteins involved in the pathogenesis of AAAs. To explore the possible pathogenetic mechanisms involved in AAA, we analyzed by proteomics modifications in plasma proteome of patients with AAA. Therefore, the present study analyzed the soluble plasma proteins using two dimensional electrophoresis (2-DE) and mass spectrometry (MS). We identified 33 protein spots, 31 of which show an up-regulation in AAA patients whilst the expression level of 2 protein spots is reduced. We confirm a number of biomarkers associated with AAA that have been previously identified by various authors. We identified a significant increase of a class of proteins such as fibrinogen, α1-antitrypsin and haptoglobin in plasma from AAA patients. The presence of these proteins in human AAA plasma may be related to the inflammatory processes active in these subjects. We have seen a negative correlation between the vitamin D-binding protein (DBP) and hemoglobin subunit β and AAA presence. DBP levels have been found to increase in AAA wall tissues by others and this discrepancy with our results could be due to the different analysis source. We wanted to analyze the factors measurable in plasma-associated rather than in tissue-associated markers because the application of circulating biomarkers in diagnostic laboratories would be relatively simple. DBP is very important for vascular remodelling and it may have an important role in the protection of vascular walls. In plasma tissue this protein reduces platelet aggregation and extends coagulation time. No one protein identified in this study has the biologic plausibility to be used singularly as a biomarker of aneurysmal disease due to inadequate specificity. The effect of using multiple biomarkers combined with clinical factors requires investigation in carefully designed population-based studies and these studies need to select the criteria of choice to define healthy controls very carefully. Clearer identification of various markers is needed, possibly using other proteomic techniques to screen for new candidates such as gel-free proteomic technology that enables us to handle larger groups of subject compared to gel-based proteomic technology. 相似文献
7.
Rusińska A Świątkowska M Koziołkiewicz W Skurzyński S Golec J Chlebna-Sokół D 《Acta biochimica Polonica》2011,58(4):553-561
The aim of the study is proteomic analysis of the plasma profile in children with recurrent bone fractures. The study involved 16 children: 6 patients with recurrent low-energy fractures and normal bone mass and 10 with osteogenesis imperfecta. In the analysis of the protein profile, the two-dimensional protein electrophoresis was used (Ettan DALT II, Amersham Bioscience). The images of protein gels were compared with controls. The protein spots with changed expression were cut from the gel and the amino acid sequence was analyzed with the mass spectrometry method (Q-Tof Premier(TM) API MASS SPECTROMETR, Waters) for protein identification. The most prevalent protein with changed expression, with respect to controls, was haptoglobin observed in 6 patients with a severe form of osteogenesis imperfecta. Increased haptoglobin concentration in these patients was confirmed by the ELISA method. Peptides corresponding to alpha-1 acid glycoprotein and serum amyloid P-component, apolipoprotein A-I, and transthyretin were detected in one, two and three children, respectively. Conclusions: 1) The results show increased haptoglobin which may be suggestive of an inflammatory component taking part in the course of osteogenesis imperfecta. 2) Further studies to explain the possible relationship of this protein with increased bone fragility are necessary. 相似文献
8.
Alonso-Orgaz S Moreno L Macaya C Rico L Mateos-Cáceres PJ Sacristán D Pérez-Vizcaíno F Segura A Tamargo J López-Farré A 《Journal of proteome research》2006,5(9):2301-2308
Proteomics is a technology to detect and identify several proteins and their isoforms in a single sample. We used proteomics to analyze modifications in the protein map of plasma after simvastatin treatment of moderate hypercholesterolemic patients. Plasma from hypercholesterolemic patients (n = 9) was compared before and after 12 weeks of simvastatin treatment (40 mg/day). Patients with similar cardiovascular risk factors were used as controls (CR group). By using two-dimensional electrophoresis and mass spectrometry, we identified the different protein isoforms. The plasma expression of three fibrinogen gamma chain isoforms (FGG) was enhanced, whereas the expression of two isoforms of the fibrinogen beta chain (FGB) was reduced in the hypercholesterolemic patients compared with the CR group. The expression of apolipoprotein A-IV and three haptoglobin isoforms was higher in hypercholesterolemic patients. Simvastatin treatment modified the plasma expression of FGG chain isoform 1, FGB chain isoforms 1 and 2, vitamin D binding protein isoform 3, apo A-IV, and haptoglobin isoform 2. The modification of FGG chain isoform 1 and FGB chain isoforms 1 and 2 was positively correlated with total plasma cholesterol level. Proteomic analysis of plasma may help to know more in depth the molecular mechanism modified by simvastatin treatment. 相似文献
9.
N. A. Pakharukova L. Kh. Pastushkova S. A. Moshkovskii I. M. Larina 《Biochemistry (Moscow) Supplemental Series B: Biomedical Chemistry》2011,5(3):203-212
The aim of this review is to analyze results of studies on characteristics of protein variability and diversity of posttranslational
modifications of proteins in healthy humans. Numerous studies have demonstrated that a proteomic profile is characterized
by significant intra- and inter-individual variability, and quite often natural (“normal”) variability of some proteins can
be comparable to changes observed in pathological processes. Results obtained by our research group have demonstrated high
intra-individual variability of serum low-molecular subproteome of healthy volunteers, certified by a special medial committee
(the coefficient of variation (CV) of 42.6%). The proteins characterized by high variability under normal conditions (e.g.
haptoglobin — 0–40 mg/ml; lysozyme — 0.01–0.1 mg/ml; C-reactive protein — 0.01–0.3 mg/ml) cannot be considered as potential
biomarkers of diseases. On the contrary, proteins and peptides characterized by insignificant dispersion in healthy population
(such as albumin ( CV = 9%); transferrin-(CV = 14%); C3c complement (CV = 17%), α-1 acid glycoprotein (CV = 21%), α-2-macroglobulin
(CV = 20%); transthyretin fragment (CV = 28.3%) and α2-HS-glycoprotein βchain (CV = 29.7%)) can provide valuable information
about the state of health. Thus, studies of plasticity in the proteomic profiles of healthy humans will help to correct reference
intervals used in clinical proteomics. 相似文献
10.
Shumei Bai Shilian Liu Xuxiao Guo Zhaoyu Qin Banqin Wang Xiaohong Li Yanjiang Qin 《Molecular biology reports》2010,37(3):1619-1625
Neuromyelitis optica (NMO) is an inflammatory demyelinating disease with generally poor prognosis that selectively targets
optic nerves and spinal cord. Although diagnostic criteria for NMO are available, there is still a need for biomarkers, predicting
disease development and progression to improve individually tailored treatment. CSF proteins were separated by two-dimensional
electrophoresis and identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF
MS). The interaction between these proteins was further analyzed by Pathway Studio software. Seven protein spots in CSF were
significantly altered in NMO patients compared with controls. Identification made by mass spectrometry revealed that the most
significant protein was haptoglobin, which was increased in the NMO gels. The subsequent ELISA test were performed to validate
it, which confirmed the results of proteomic analysis. Protein network was built, which showed some biological interactions
among the seven proteins. These results support a correlation between the level of haptoglobin and NMO. Haptoglobin may be
a potential useful biomarker for diagnosis or a medicine target for treatment of NMO. 相似文献
11.
Yasmin Ahmad Narendra K. Sharma Iti Garg Mohammad Faiz Ahmad Manish Sharma Kalpana Bhargava 《PloS one》2013,8(7)
Adaptation to hypobaric hypoxia is required by animals and human in several physiological and pathological situations. Hypobaric hypoxia is a pathophysiological condition triggering redox status disturbances of cell organization leading, via oxidative stress, to proteins, lipids, and DNA damage. Identifying the molecular variables playing key roles in this process would be of paramount importance to shed light on the mechanisms known to counteract the negative effects of oxygen lack. To obtain a molecular signature, changes in the plasma proteome were studied by using proteomic approach. To enrich the low-abundance proteins in human plasma, two highly abundant proteins, albumin and IgG, were first removed. By comparing the plasma proteins of high altitude natives with those of a normal control group, several proteins with a significant alteration were found. The up-regulated proteins were identified as vitamin D-binding protein, hemopexin, alpha-1–antitrypsin, haptoglobin β-chain, apolipoprotein A1, transthyretin and hemoglobin beta chain. The down-regulated proteins were transferrin, complement C3, serum amyloid, complement component 4A and plasma retinol binding protein. Among these proteins, the alterations of transthyretin and transferrin were further confirmed by ELISA and Western blotting analysis. Since all the up- and down- regulated proteins identified above are well-known inflammation inhibitors and play a positive anti-inflammatory role, these results show that there is some adaptive mechanism that sustains the inflammation balance in high altitude natives exposed to hypobaric hypoxia. 相似文献
12.
Marta Piroddi Angelo Palmese Francesca Pilolli Angela Amoresano Piero Pucci Claudio Ronco Francesco Galli 《Amino acids》2011,40(2):653-667
3′-nitrotyrosine (3NT) is a post-translational modification (PTM) of body fluids and tissues that is sustained by chronic
inflammation and oxidative stress, two main clinical traits of chronic kidney disease (CKD). Despite this background, protein
targets and their differential susceptibility to in vivo nitration remain almost completely unexplored in CKD. This study
reports a first investigation of plasma nitroproteome in these patients, carried out by both immunorecognition and LC-MS/MS
techniques. Plasma proteins of chronic and end-stage KD patients showed a higher burden of nitration than in healthy controls,
but main nitration targets appeared to be the same in these populations. Immunoblotting data showed that uremic albumin is
largely represented in the uremic nitroproteome together with fibrinogen chains (A, B and C), transferrin, α1-antitrypsin,
complement factor D, haptoglobin, and IgG light and heavy chains. However, immunopurification and affinity chromatography
experiments demonstrated that the relative content of 3NT on the albumin molecule was very low when compared with that of
the remaining plasma proteins. The uremic nitroproteome was investigated using also plasma proteins obtained by in vivo ultrafiltration
from patients treated with protein leaking or standard high-flux hemodialyzers. The study of these samples revealed the possibility
to selectively remove protein nitration products during hemodialysis. Identification of intramolecular sites of nitration
was preliminarily obtained in IgG chains isolated by 2D PAGE and assessed by bidimensional tandem mass spectrometry after
chemoselective tagging. Further studies are needed to confirm at the molecular level the presence of nitrated Tyr residues
in other proteins tentatively identified as nitration targets in this study and to explore the biological meaning of such
a selective modification of plasma proteins by reactive nitrogen species in uremia and dialysis patients. 相似文献
13.
Peili Zhang Zhanquan Li Faman Yang Linhua Ji Yingzhong Yang Chuanchuan Liu Huihui Liu Jie Ma Jie Liu Zhancui Dang Shengyan Wang Rili Ge Sen Cui 《Bioscience reports》2021,41(1)
Chronic mountain sickness (CMS) is a progressive incapacitating syndrome induced by lifelong exposure to hypoxia. In the present study, proteomic analysis was used to identify the differentially expressed proteins (DEPs) and then evaluate the potential plasma biomarkers between CMS and non-CMS groups. A total of 145 DEPs were detected in CMS Han Chinese people who live in the plateau (CMS-HPu), among which 89 were significantly up-regulated and 56 were significantly down-regulated. GO enrichment analysis showed that various biological processes were enriched, including the hydrogen peroxide metabolic/catabolic process, reactive oxygen species (ROS) metabolic, and acute inflammatory response. Protein–protein interaction analysis showed that antioxidant activity, the hydrogen peroxide catabolic process and peroxidase activity were primarily mapped in interaction proteins. Nine modules showed significantly clustering based on WGCNA analysis, with two being the most significant, and GO analysis showed that proteins of both modules were primarily enriched in oxidative stress-related biological processes. Four DEPs increased in CMS patients were evaluated as the candidate biomarkers, and three showed significant AUC: hemoglobin β chain (HB-β), thioredoxin-1 (TRX1), and phosphoglycerate kinase 1 (PGK1). The present study provides insights into the pathogenesis of CMS and further evaluates the potentially biomarkers for its prevention and treatment of it. 相似文献
14.
Madhulika B. Gupta Maxim D. Seferovic Suya Liu Robert J. Gratton Amanda Doherty-Kirby Gilles A. Lajoie Victor K. M. Han 《Clinical proteomics》2006,2(3-4):169-184
Fetal growth restriction (FGR) affects 3–5% of pregnancies and is associated with increased perinatal morbidity and mortality.
Currently, there is no reliable biochemical test to differentiate a pathological FGR from a nonpathological one. The objective
of this study was to screen whole maternal plasma to identify differentially expressed relatively abundant proteins associated
with FGR. We analyzed maternal plasma from FGR (n=28) and healthy (n=22) pregnancies using two-dimensional gel electrophoresis (2D-GE) followed by software image analysis. Three spots with molecular
weight (Mr) 18 kDa corresponding to haptoglobin (hp) α2, as identified by LC-MS/MS and immunoblotting, showed differential expression
patterns in FGR. The distribution of hp α2 variants in maternal plasma samples showed the hp α2 variant 1 was low in 72% of
FGR, medium in 16%, whereas high in 12%. In comparison, hp α2 variant 1 was high in (41%) of controls, medium in 41%, and
low in 18% of cases. Based on the software image analysis, the mean spot volume for hp α2 variant 1 was 0.12 (SD=0.18) for
FGR compared to 0.26 (SD=0.19) for control (p=0.006). Given that hp turnover is indicative of its maturation process and is traceable in plasma by its dominant/suppressed
variants, we propose that hp α2 is an important potential target for evaluation of its clinical and pathophysiological role
and as a diagnostic biomarker in FGR. 相似文献
15.
Inflammation inhibitors were remarkably up-regulated in plasma of severe acute respiratory syndrome patients at progressive phase 总被引:3,自引:0,他引:3
Wan J Sun W Li X Ying W Dai J Kuai X Wei H Gao X Zhu Y Jiang Y Qian X He F 《Proteomics》2006,6(9):2886-2894
Severe acute respiratory syndrome (SARS) is a severe infectious disease that has affected many countries and regions since 2002. A novel member of the coronavirus, SARS-CoV, has been identified as the causative agent. However, the pathogenesis of SARS is still elusive. In this study, we used 2-D DIGE and MS to analyze the protein profiles of plasma from SARS patients, in the search for proteomic alterations associated with the disease progression, which could provide some clues to the pathogenesis. To enrich the low-abundance proteins in human plasma, two highly abundant proteins, albumin and IgG, were first removed. By comparing the plasma proteins of SARS patients with those of a normal control group, several proteins with a significant alteration were found. The up-regulated proteins were identified as alpha-1 acid glycoprotein, haptoglobin, alpha-1 anti-chymotrypsin and fetuin. The down-regulated proteins were apolipoprotein A-I, transferrin and transthyretin. Most of the proteins showed significant changes (up- or down-regulated) in the progressive phase of disease, and there was a trend back to normal level during the convalescent phase. Among these proteins, the alterations of fetuin and anti-chymotrypsin were further confirmed by Western blotting. Since all the up-regulated proteins identified above are well-known inflammation inhibitors, these results strongly suggest that the body starts inflammation inhibition to sustain the inflammatory response balance in the progression of SARS. 相似文献
16.
Raghav SK Gupta B Agrawal C Saroha A Das RH Chaturvedi VP Das HR 《Glycoconjugate journal》2006,23(3-4):167-173
Altered glycosylation of plasma proteins has been directly implicated in the pathogenesis of rheumatoid arthritis (RA). The
present study investigated the changes in the Concanavalin-A (Con-A)-bound plasma proteins in the RA patients in comparison to that of the healthy controls. Two proteins (MW ∼32 kDa
and ∼62 kDa) showed an alteration in expression while an altered monosaccharide profile (high mannose) was observed in the
∼62 kDa protein in the samples collected from RA patients. The 2-dimensional polyacrylamide gel electrophoresis analysis of
the Con-A-bound plasma samples showed a large number of protein spots, a few of which were differentially expressed in the
RA patients. Some unidentified proteins were detected in the RA patients which were absent in the control samples. The present
study, therefore, enunciates the role of carbohydrates as well as that of the acute phase response in the disease pathogenesis. 相似文献
17.
Linke T Doraiswamy S Harrison EH 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2007,849(1-2):273-281
The proteomic analysis of plasma and serum samples represents a formidable challenge due to the presence of a few highly abundant proteins such as albumin and immunoglobulins. Detection of low abundance protein biomarkers requires therefore either the specific depletion of high abundance proteins with immunoaffinity columns and/or optimized protein fractionation methods based on charge, size or hydrophobicity. Here we describe the depletion of seven abundant rat plasma proteins with an immunoaffinity column with coupled antibodies directed against albumin, IgG, transferrin, IgM, haptoglobin, fibrinogen and alpha1-anti-trypsin. The IgY-R7-LC2 (Beckman Coulter) column showed high specificity for the targeted proteins and was able to efficiently remove most of the albumin, IgG and transferrin from rat plasma samples as judged by Western blot analysis. Depleted rat plasma protein samples were analyzed by SELDI-TOF MS, 2D SDS-PAGE and 2D-LC and compared to non-depleted plasma samples as well as to the abundant protein fraction that was eluted from the immunoaffinity column. Analysis of the depleted plasma protein fraction revealed improved signal to noise ratios, regardless of which proteomic method was applied. However, only a small number of new proteins were observed in the depleted protein fraction. Immunoaffinity depletion of abundant plasma proteins results in the significant dilution of the original sample which complicates subsequent analysis. Most proteomic approaches require specialized sample preparation procedures during which significant losses of less abundant proteins and potential biomarkers can occur. Even though abundant protein depletion reduces the dynamic range of the plasma proteome by about 2-3 orders of magnitude, the difference between medium-abundant and low abundant plasma proteins is still in the range of 7-8 orders of magnitude and beyond the dynamic range of current proteomic technologies. Thus, exploring the plasma proteome in greater detail remains a daunting task. 相似文献
18.
Xiaoping Ao Lujun Zhao Mary A Davis David M Lubman Theodore S Lawrence Feng-Ming Kong 《Journal of hematology & oncology》2009,2(1):1-12
We studied serum proteomic profiling in patients with graft versus host disease (GVHD) after allogeneic hematopoietic cell transplantation (allo-HCT) by two-dimensional gel electrophoresis (2-DE) and mass spectrometry analysis. The expression of a group of proteins, haptoglobin (Hp), alpha-1-antitrypsin, apolipoprotein A-IV, serum paraoxonase and Zn-alpha-glycoprotein were increased and the proteins, clusterin precursor, alpha-2-macroglobulin, serum amyloid protein precursor, sex hormone-binding globulin, serotransferrin and complement C4 were decreased in patients with extensive chronic GVHD (cGVHD). Serum haptoglobin (Hp) levels in patients with cGVHD were demonstrated to be statistically higher than in patients without cGVHD and normal controls (p < 0.01). We used immunoblotting and PCR in combination with 2-DE gel image analysis to determine Hp polymorphisms in 25 allo-HCT patients and 16 normal donors. The results demonstrate that patients with cGVHD had a higher incidence of HP 2-2 phenotype (43.8%), in comparison to the patients without cGVHD (0%) and normal donors (18.7%), suggesting the possibility that specific Hp polymorphism may play a role in the development of cGVHD after allo-HCT. In this study, quantitative serum Hp levels were shown to be related to cGVHD development. Further, the data suggest the possibility that specific Hp polymorphisms may be associated with cGVHD development and warrant further investigation. 相似文献
19.
Scoppetta F Tartaglia M Renzone G Avellini L Gaiti A Scaloni A Chiaradia E 《Journal of Proteomics》2012,75(14):4494-4504
Physical exercise induces various stress responses and metabolic adaptations that have not yet been completely elucidated. Novel biomarkers are needed in sport veterinary medicine to monitor training levels and to detect subclinical conditions that can develop into exercise-related diseases. In this study, protein modifications in horse plasma induced by prolonged, aerobic physical exercise were investigated by using a proteomic approach based on 2-DE and combined mass spectrometry procedures. Thirty-eight protein spots, associated with expression products of 13 genes, showed significant quantitative changes; spots identified as membrane Cu amine oxidase, α-1 antitrypsin, α-1 antitrypsin-related protein, caeruloplasmin, α-2 macroglobulin and complement factor C4 were augmented in relative abundance after the race, while haptoglobin β chain, apolipoprotein A-I, transthyretin, retinol binding protein 4, fibrinogen γ chain, complement factor B and albumin fragments were reduced. These results indicate that prolonged physical exercise affects plasma proteins involved in pathways related to inflammation, coagulation, immune modulation, oxidant/antioxidant activity and cellular and vascular damage, with consequent effects on whole horse metabolism. 相似文献
20.
Yang YR Liu SL Qin ZY Liu FJ Qin YJ Bai SM Chen ZY 《Cellular and molecular neurobiology》2008,28(5):737-744
To better understand the pathophysiologic mechanisms underlying Guillain-Barré syndrome (GBS), Comparative proteomic analysis of cerebrospinal fluid (CSF) between patients with GBS (the experiment group) and control subjects suffering from other neurological disorders (the control group) was carried out using two-dimensional gel electrophoresis (2-DE) technique, in combination with matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) and database searching to determine abnormal CSF proteins in GBS patients. Image analysis of 2-DE gels silver stained revealed that 10 protein spots showed significant differential expression between the two groups of CSF samples. The expression of cystatin C, transthyretin, apolipoprotein E and heat shock protein 70 were decreased. However, haptoglobin, alpha-1-antitrypsin, apolipoprotein A-IV and neurofilaments were elevated. The subsequent ELISA measured the concentration of cystatin C and confirmed the result of the proteomic analysis. These identified proteins may be involved in the pathophysiological process of GBS and call for further studying the role of these proteins in the pathogenesis of the disease. 相似文献