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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.
Summary. Ten years after the establishment of the term proteome, the science surrounding it has yet to fulfill its potential. While
a host of technologies have generated lists of protein names, there are only a few reported studies that have examined the
individual proteins at the covalent chemical level defined as protein species in 1997 and their function. In the current study,
we demonstrate that this is possible with two-dimensional gel electrophoresis (2-DE) and mass spectrometry by presenting clear
evidence of in vivo N-terminal alpha A crystallin truncation and relating this newly detected protein species to alpha crystallin
activity regulation by protease cleavage in the healthy young murine lens. We assess the present state of technology and suggest
a shift in resources and paradigm for the routine attainment of the protein species level in proteomics. 相似文献
3.
Summary. The eye lens is a fascinating organ as it is in essence living transparent matter. Lenticular transparency is achieved through
the peculiarities of lens morphology, a semi-apoptotic process where cells elongate and loose their organelles and the precise
molecular arrangement of the bulk of soluble lenticular proteins, the crystallins. The 16 crystallins ubiquitous in mammals
and their modifications have been extensively characterized by 2-DE, liquid chromatography, mass spectrometry and other protein
analysis techniques. The various solubility dependant fractions as well as subproteomes of lenticular morphological sections
have also been explored in detail. Extensive post translational modification of the crystallins is encountered throughout
the lens as a result of ageing and disease resulting in a vast number of protein species. Proteomics methodology is therefore
ideal to further comprehensive understanding of this organ and the factors involved in cataractogenesis. 相似文献
4.
Mass-spectrometrical analysis of proteins encoded on chromosome 21 in human fetal brain 总被引:1,自引:0,他引:1
Summary. Overexpression of chromosome 21 genes is directly or indirectly responsible for the Down syndrome phenotype. In order to analyse
chromosome 21 gene products (Chr21Ps), we extracted proteins from fetal human brain cortex and applied an ultracentrifugal
and chromatographic prefractionation principle followed by two-dimensional gel electrophoresis (2-DE) and mass-spectrometrical
analysis using high-throughput automated MALDI-TOF/TOF. Nine Chr21Ps were identified: pyridoxal kinase; superoxide dismutase
[Cu/Zn] 1; carbonyl reductase 1; ES1 protein homolog, mitochondrial [Precursor]; cystathionine-beta-synthetase; T-complex
protein 1, theta subunit; cystatin B; 6-phosphofructokinase; glycinamide ribonucleotide synthetase. Mass-spectrometric characterisation
of Chr21Ps following separation in 2-DE gels is a useful tool for the analysis of these structures in brain, independent of
antibody availability and specificity. 相似文献
5.
Aberrant expression of cytoskeleton proteins in hippocampus from patients with mesial temporal lobe epilepsy 总被引:8,自引:0,他引:8
Summary. Mesial temporal lobe epilepsy (MTLE), the most common form of epilepsy, is characterised by cytoarchitectural abnormalities
including neuronal cell loss and reactive gliosis in hippocampus. Determination of aberrant cytoskeleton protein expression
by proteomics techniques may help to understand pathomechanism that is still elusive. We searched for differential expression
of hippocampal proteins by an analytical method based on two-dimensional gel electrophoresis (2-DE) coupled with mass spectrometry
unambiguously identifying 77 proteins analysed in eight control and eight MTLE hippocampi. Proteins were quantified and we
observed 18 proteins that were altered in MTLE. Cytoskeleton proteins tubulin α-1 chain, β-tubulin, profilin II, neuronal
tropomodulin were significantly reduced and one actin spot was missing, whereas ezrin and vinculin were significantly increased
in MTLE. Proteins of several classes as e.g. antioxidant proteins (peroxiredoxins 3 and 6), chaperons (T-complex protein 1-α,
stress-induced-phosphoprotein 1), signaling protein MAP kinase kinase 1, synaptosomal proteins (synaptotagmin I, α-synuclein),
NAD-dependent deacetylase sirtuin-2 and 26S protease regulatory subunit 7 protein, neuronal-specific septin 3 were altered
in MTLE. Taken together, the findings may represent or lead to cytoskeletal impairment; aberrant antioxidant proteins, chaperons,
MAP kinase kinase 1 and NAD-dependent deacetylase sirtuin-2 may have been involved in pathogenetic mechanisms and altered
synaptosomal protein expression possibly reflects synaptic impairment in MTLE.
J. W. Yang and T. Czech have equally contributed to the paper. 相似文献
6.
Carbonaro M 《Amino acids》2006,31(4):485-488
Summary. Two-dimensional electrophoresis (2-DE) was used for tracing in vivo gastrointestinal digestion of milk proteins in a rapid
model system with rats. Contents of stomach and small intestine from digestion trials with rats given a single dose of milk
powder were recovered after 1 hour. They were then subjected to 2-DE (IEF and SDS-PAGE). 2-DE showed undigested proteins in
a MW range 13.0–66.0 kDa in stomach and 13.0–25.0 kDa in the small intestine, thus indicating that milk proteins are slowly
digested. This approach may shed light on pattern of protein digestion and mechanism of amino acid and peptide assimilation. 相似文献
7.
Summary. In plasma and serum, the presence of high-abundance proteins can overwhelm the signals of low-abundance proteins, which then
become undetectable either by two-dimensional gels or chromatographic techniques. Therefore, depletion of abundant proteins
is a prerequisite to detect low-abundance components. Furthermore, the regeneration of pre-purification tools could be money-saving.
We applied an affinity chromatography kit to remove albumin and the immunoglobulin chains from plasma and propose a simple
and effective technical procedure for the regeneration of these affinity columns.
The first two authors have equally contributed to this work.
Authors’ address: Prof. Paolo Tessari, Dipartimento di Medicina Clinica e Sperimentale, Università di Padova, via Giustiniani
2, 35128 Padova, Italy 相似文献
8.
Update and challenges on proteomics in rice 总被引:4,自引:0,他引:4
Rice is not only an important agricultural resource but also a model plant for biological research. Our previous review highlighted different aspects of the construction of rice proteome database, cataloguing rice proteins of different tissues and organelle, differential proteomics using 2-DE and functional characterization of some of the proteins identified (Komatsu, S., Tanaka, N., Proteomics 2005, 5, 938-949). In this review, the powerfulness and weaknesses of proteomic technologies as a whole and limitations of the currently used techniques in rice proteomics are discussed. The information obtained from these techniques regarding proteins modification, protein-protein interaction and the development of new methods for differential proteomics will aid in deciphering more precisely the functions of known and/or unknown proteins in rice. 相似文献
9.
Keck ME 《Amino acids》2006,31(3):241-250
Summary. Affective disorders tend to be chronic and life-threatening diseases: suicide is estimated to be the cause of death in 10–15%
of individuals with major depressive disorders. Major depression is one of the most prevalent and costly brain diseases with
up to 20% of the worldwide population suffering from moderate to severe forms of the disease. Only 50% of individuals with
depression show full remission in response to currently available antidepressant drug therapies which are based on serendipitous
discoveries made in the 1950s. Previously underestimated, other severe depression-associated deleterious health-related effects
have increasingly been recognized. Epidemiological studies have provided substantial evidence that patients with depression
have a 2–4-fold increased risk both of developing cardiovascular disease and of mortality after experiencing a myocardial
infarction. The majority of patients suffering from affective disorders have measurable shifts in their stress hormone regulation
as reflected by elevated secretion of central and peripheral stress hormones or by altered hormonal responses to neuroendocrine
challenge tests. In recent years, these alterations have increasingly been translated into testable hypotheses addressing
the pathogenesis of illness. Refined molecular technologies and the creation of genetically engineered mice have allowed to
specifically target individual genes involved in regulation of corticotropin releasing factor (CRF) and vasopressin (AVP)
system elements. The cumulative evidence makes a strong case implicating dysfunction of these systems in the etiology and
pathogenesis of depression and pathological anxiety. Translation of these advances into novel therapeutic strategies has already
been started. 相似文献
10.
Summary. Pseudomonas sp. strain phDV1, being a phenol degrading bacterium, has been found to utilize phenol as sole carbon source via the meta pathway. Blue native polyacrylamide gel electrophoresis (BN-PAGE) is widely used for the analysis of oligomeric state and
molecular mass non-dissociated protein complexes. In this study, a number of proteomic techniques were used to investigate
the oligomeric state enzymes involved in the aromatic degradation pathway. In particular, the Pseudomonas sp. strain phDV1 proteome was monitored under two different growth substrate conditions, using glucose or phenol as sole
carbon source. The protein complexes map was compared by BN-PAGE after fractionation by sucrose density centrifugation of
the cell extracts. Multiple differences were detected. Further, analysis and identification of the subunit composition of
these complexes was carried out using MALDI-TOF MS, allowing the identification of 49 proteins. Additionally, functional information
regarding protein–protein interactions was assembled, by coupling 2-D BN-PAGE with MALDI-TOF MS. Application of this functional
proteomics method resulted in an higher number of the identified proteins. 相似文献
11.
Cartilage is a highly specialized load-bearing tissue with a small number of cells and a high proportion of extracellular matrix (ECM). The abundance of heavily sulfated proteoglycans and a poorly soluble collagenous ECM presents a major technical challenge to 2-DE. Here we report proteomic analysis of mouse growth plate cartilage using novel methodology for tissue dissection and sample prefractionation. We have successfully resolved cartilage tissue extracts by 2-DE for the first time and identified cartilage ECM proteins by Western blotting and MS/MS. 相似文献
12.
Summary. No systematic approach to detect expression of differentiation-related elements was published so far. The undifferentiated
N1E-115 neuroblastoma cell line was switched into a neuronal phenotype by DMSO treatment and used for proteomic experiments.
We used two-dimensional gel electrophoresis followed by unambiguous mass spectrometrical identification of proteins to generate
a map of cytoskeleton proteins (CPs), i.e., to search for differentiation-related structures. Alpha-actin, actin-like protein
6A, gamma-tubulin complex component 2, tubulin alpha 3/alpha 7, CLIP associating protein 2, B4 integrin interactor homolog
were detectable in the undifferentiated cell line exclusively and neuron-specific CPs drebrin and presynaptic density protein
95, actin-related protein 2/3, alpha and beta-centractin, PDZ-domain actin binding protein, actinin alpha 1, profilin II,
ezrin, coactosin-like protein, transgelin 2, myosin light polypeptide 6, tubulin alpha 2, 6 and 7, beta tubulin (94% similar
with tubulin beta-2), tubulin beta 3, tubulin tyrosine ligase-like protein 1, lamin B1 and keratin 20 were observed in the
differentiated cell line only. We herein identified differentiation-related expressional patterns thus providing new evidence
for the role of CPs in the process of neuronal differentiation. 相似文献
13.
Two-dimensional electrophoresis (2-DE) is one of the most commonly used techniques in proteomic investigations. However, due to the complex interplay of incidence including significant biological sample variations, lengthy steps involved in performing 2-DE as well as exposure time with silver staining, it is sometimes difficult to differentiate authentic differences caused by drug treatment with those artifacts caused by sample variations, running conditions of 2-DE as well as treatment time in silver staining etc. If we can compare pooled samples of control and treatment groups run in a single gel and stained together, we would be more comfortable with our findings. We propose here a low cost and highly effective method for locating differentially expressed proteins before and after drug treatment. This "two-in-one gel" technique might partially solve the problems mentioned above. 相似文献
14.
Summary. In the postgenomic era new technologies are emerging for global analysis of protein function. The introduction of active site-directed
chemical probes for enzymatic activity profiling in complex mixtures, known as activity-based proteomics has greatly accelerated
functional annotation of proteins. Here we review probe design for different enzyme classes including serine hydrolases, cysteine
proteases, tyrosine phosphatases, glycosidases, and others. These probes are usually detected by their fluorescent, radioactive
or affinity tags and their protein targets are analyzed using established proteomics techniques. Recent developments, such
as the design of probes for in vivo analysis of proteomes, as well as microarray technologies for higher throughput screenings
of protein specificity and the application of activity-based probes for drug screening are highlighted. We focus on biological
applications of activity-based probes for target and inhibitor discovery and discuss challenges for future development of
this field. 相似文献
15.
蛋白质组学研究中的新技术 总被引:1,自引:0,他引:1
随着后基因组时代的到来,蛋白质组研究日益受到密切关注。蛋白质组学的发展需要有先进的技术平台作支撑。除了蛋白质组研究中的三大核心技术-双向电泳、质谱和生物信息学技术,近几年又不断涌现出许多新技术。本文对同位素标记亲和标签、酵母双杂交、多维色谱—质谱联用等新技术作一概述。 相似文献
16.
Aberrant cytosolic acyl-CoA thioester hydrolase in hippocampus of patients with mesial temporal lobe epilepsy 总被引:2,自引:0,他引:2
Summary. A series of enzyme alterations has been shown to be associated with several forms of epilepsy, in mesial temporal lobe epilepsy (MTLE), however, information is limited. It was therefore the aim of the study to determine brain enzyme protein expression using a proteomic screening approach. Hippocampi of controls and patients with drug-resistant MTLE were used for evaluation of protein expression. We applied two-dimensional electrophoresis (2-DE) with mass spectrometrical identification and immunoblotting. 2-DE revealed a remarkably decreased spot identified as cytosolic acyl-CoA thioester hydrolase (BACH; EC 3.1.2.2) in patients with MTLE. Western blotting showed absence of bands at 37kDa in MTLEs using an antibody against mouse BACH and at 140kDa in MTLEs using anti-rat BACH. This study demonstrates that BACHs were deranged in hippocampus of MTLE patients. This finding may well contribute to the understanding of the still elusive pathomechanisms involved in MTLE.J. W. Yang and T. Czech have equally contributed to the paper. 相似文献
17.
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
18.
Stout JR Cramer JT Zoeller RF Torok D Costa P Hoffman JR Harris RC O'Kroy J 《Amino acids》2007,32(3):381-386
Summary. This study examined the effects of 28 days of β-alanine supplementation on the physical working capacity at fatigue threshold
(PWCFT), ventilatory threshold (VT), maximal oxygen consumption (
O2-MAX), and time-to-exhaustion (TTE) in women. Twenty-two women (age ± SD 27.4 ± 6.1 yrs) participated and were randomly assigned
to either the β-alanine (CarnoSyn™) or Placebo (PL) group. Before (pre) and after (post) the supplementation period, participants
performed a continuous, incremental cycle ergometry test to exhaustion to determine the PWCFT, VT,
O2-MAX, and TTE. There was a 13.9, 12.6 and 2.5% increase (p < 0.05) in VT, PWCFT, and TTE, respectively, for the β-alanine group, with no changes in the PL (p > 0.05). There were no changes for
O2-MAX (p > 0.05) in either group. Results of this study indicate that β-alanine supplementation delays the onset of neuromuscular
fatigue (PWCFT) and the ventilatory threshold (VT) at submaximal workloads, and increase in TTE during maximal cycle ergometry performance.
However, β-alanine supplementation did not affect maximal aerobic power (
O2-MAX). In conclusion, β-alanine supplementation appears to improve submaximal cycle ergometry performance and TTE in young women,
perhaps as a result of an increased buffering capacity due to elevated muscle carnosine concentrations. 相似文献
19.
20.
Proteomic analysis of cartilage proteins 总被引:1,自引:0,他引:1
While the analysis of the cartilage proteome is important for our comprehensive understanding of the development and disease of this important tissue, several unique features of cartilage present some technical obstacles. Firstly, cartilage is difficult to obtain in adequate quantities for many protein analyses, especially from mice which are otherwise powerful experimental models. Furthermore, the cartilage extracellular matrix contains an insoluble network of collagen II-containing fibrils that are integrated within an abundant anionic network of aggrecan and hyaluronan aggregates. These interacting networks provide a structural scaffold for the covalent and non-covalent attachment of other proteins and glycoproteins. Consequently, proteomic analysis of cartilage requires extraction of proteins with chaotropic agents to achieve and significant protein solubilization. Finally, isolated chondrocytes are phenotypically unstable, which requires rapid isolation of cells or the use of specific culture conditions. Despite these problems, recent improvements in the sensitivity and reproducibility of two-dimensional electrophoresis (2-DE) and tandem mass spectrometry (MS/MS) techniques, combined with improved tissue preparation and sample pre-fractionation approaches, have made the proteomic characterization of cartilage tissues possible. Here we review the approaches that have been used and describe in detail protocols for the proteomic analysis of cartilage tissues and cells. 相似文献