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1.
Profiling of the secreted proteins during 3T3-L1 adipocyte differentiation leads to the identification of novel adipokines 总被引:3,自引:0,他引:3
Wang P Mariman E Keijer J Bouwman F Noben JP Robben J Renes J 《Cellular and molecular life sciences : CMLS》2004,61(18):2405-2417
Adipose tissue is an endocrine organ capable of secreting a number of adipokines with a role in the regulation of
adipose tissue and whole-body metabolism. We used two-dimensional gel electrophoresis combined with mass spectrometry to
profile the secreted proteins from (pre)adipocytes. The culture medium of 3T3-L1 cells during adipocyte differentiation
was screened, and 41 proteins that responded to blocking of secretion by 20°C treatment and/or brefeldin A treatment
were identified. Prohibitin, stress-70 protein, and adhesion-regulating molecule 1 are reported for the first time as
secreted proteins. In addition, procollagen C-proteinase enhancer protein, galectin-1, cyclophilin A and C, and SF20/IL-25
are newly identified as adipocyte secreted factors. Secretion profiles indicated a dynamic environment including an
actively remodeling extracellular matrix and several factors involved in growth regulation.Received 15 June 2004; received after revision 26 July 2004; accepted 2 August 2004 相似文献
2.
Protein kinase-dependent overexpression of the nuclear protein pirin in c-JUN and RAS transformed fibroblasts 总被引:1,自引:0,他引:1
Bergman AC Alaiya AA Wendler W Binetruy B Shoshan M Sakaguchi K Bergman T Kronenwett U Auer G Appella E Jörnvall H Linder S 《Cellular and molecular life sciences : CMLS》1999,55(3):467-471
Signalling via the protein kinase Raf-MEK-ERK pathway is of major importance for transformation by oncogenes. To identify
genes affected by inhibition of this pathway, c-JUN transformed rat fibroblasts were treated with a MEK1 inhibitor (PD98059) and subjected to two-dimensional gel electrophoresis
after cell lysis. Gene products with expression influenced by MEK1 inhibition were determined by mass spectrometry of fragments
from in-gel tryptic digestions. The expression of pirin, a nuclear factor I-interacting protein, was lowered after inhibition
of MEK1. Western blot analysis revealed increased expression of pirin in RAS and c-JUN transformed cells in the absence of PD98059. Inhibition of MEK1 also led to reduced expression of α-enolase, phosphoglycerate kinase, elongation factor 2 and heterogeneous nuclear ribonucleoprotein A3, the latter two being
detected as truncated proteins. In contrast, the level of ornithine aminotransferase was increased. We conclude that inhibition
of MEK1 results in major alterations of protein expression in c-JUN transformed cells, suggesting that this pathway is important for oncogene-induced phenotypic changes.
Received 30 December 1998; accepted 12 January 1999 相似文献
3.
Hirschberg D Cederlund E Crosas B Jonsson A Tryggvason S Farrés J Parés X Bergman T Jörnvall H 《Cellular and molecular life sciences : CMLS》2001,58(9):1323-1326
A recent finding of a novel class of retinol-active alcohol dehydrogenase (ADH) in frog prompted analysis of this activity in other vertebrate forms. Surprisingly, yet another and still more unrelated ADH was identified in chicken tissues. It was found to be a member of the aldo-keto reductase (AKR) enzyme family, not previously known as an ADH in vertebrates. Its terminal blocking group and the N-terminal segment, not assigned by protein and cDNA structure analysis, were determined by electrospray tandem mass spectrometry after protein isolation by two-dimensional gel electrophoresis. The N terminus is Acetyl-Ala- and the N-terminal segment contains two consecutive Asn residues. The results establish the new ADH enzyme of the AKR family and show the usefulness of combined gel separation and mass spectrometry in enzyme-characterization. 相似文献
4.
Roblick UJ Hirschberg D Habermann JK Palmberg C Becker S Krüger S Gustafsson M Bruch HP Franzén B Ried T Bergmann T Auer G Jörnvall H 《Cellular and molecular life sciences : CMLS》2004,61(10):1246-1255
Changes in the proteome of colon mucosal cells accompany the transition from normal mucosa via adenoma and invasive cancer to metastatic disease. Samples from 15 patients with sporadic sigmoid cancers were analyzed. Proteins were separated by two-dimensional gel electrophoresis. Relative differences in expression levels between normal tissue, adenoma, carcinoma and metastasis were evaluated in both intra- and inter-patient comparisons. Up- and down-regulated proteins (<twofold) during development to cancer or metastasis were excised and submitted to peptide mass fingerprinting and MS/MS sequence analysis, facilitated by the use of a compact disc workstation. In total, 112 protein spots were found to be differentially regulated, of which 72 were determined as to protein identity, 46 being up-regulated toward the progression of cancer, and 26 down-regulated. Several of the identifications correlate with proteins of the cell cycle, cytoskeleton or metabolic pathways. The pattern changes now identified have the potential for design of marker panels for assistance in diagnostics and therapeutic strategies in colorectal cancer.Received 2 February 2004; received after revision 16 March 2004; accepted 18 March 2004 相似文献
5.
Jägerbrink T Lexander H Palmberg C Shafqat J Sharoyko V Berggren PO Efendic S Zaitsev S Jörnvall H 《Cellular and molecular life sciences : CMLS》2007,64(10):1310-1316
The effects of an imidazoline compound (BL11282) on protein expression in rat pancreatic islets were investigated with a proteomic
approach. The compound increases insulin release selectively at high glucose concentrations and is therefore of interest in
type 2 diabetes. Whole cell extracts from isolated drug-treated and native pancreatic rat islets were compared after separation
by 2-D gel electrophoresis. Differentially expressed proteins were identified by mass spectrometry; 15 proteins were selectively
up-regulated and 7 selectively down-regulated in drug-treated islets. Of special interest among the differentially expressed
proteins are those involved in protein folding (Hsp60, protein disulfide isomerase, calreticulin), Ca2+ binding (calgizzarin, calcyclin and annexin I) and metabolism or signalling (pyruvate kinase, alpha enolase and protein kinase
C inhibitor 1).
Received 19 March 2007; received after revision 11 April 2007; accepted 11 April 2007 相似文献
6.
Renes J Bouwman F Noben JP Evelo C Robben J Mariman E 《Cellular and molecular life sciences : CMLS》2005,62(4):492-503
The increased incidence of obesity and related disorders in Western societies requires a thorough understanding of the adipogenic process. Data at the protein level of this process are scarce. Therefore we performed a proteome analysis of differentiating and starving 3T3-L1 cells using two-dimensional gel electrophoresis combined with mass spectrometry. Effects of different starvation conditions were examined by subjecting 3T3-L1 adipocytes to caloric restriction, either in the absence or the presence of the lipolysis inducer tumor necrosis factor-. Ninety-three differentially expressed proteins were found during differentiation and starvation of 3T3-L1 cells, 50 of which were identified. GenMAPP/MAPP-finder software revealed a non-reciprocal regulation of the glycolytic pathway during 3T3-L1 differentiation followed by starvation. Furthermore, proteins involved in growth regulation, cytoskeletal rearrangements and protein modification, 16 of which have not been described before in 3T3-L1 cells, were identified. In conclusion, our data provide valuable information for further understanding of the adipogenic process.Received 9 November 2004; received after revision 21 December 2004; accepted 28 December 2004 相似文献
7.
Tibaldi E Arrigoni G Brunati AM James P Pinna LA 《Cellular and molecular life sciences : CMLS》2006,63(3):378-389
In an attempt to gain information about the identity of the Golgi apparatus casein kinase(s) (G-CK), responsible for the phosphorylation
of caseins in lactating mammary gland, the proteins present in fractions enriched in G-CK activity eluted from DEAE-Sepharose
and heparin-Sepharose columns were resolved by two-dimensional electrophoresis and analyzed by mass spectrometry. This led
to the identification of 47 proteins altogether, none of which is a bona fide protein kinase. At least 9 of the identified
proteins however, are readily phosphorylated by co-purifying G-CK activity, and 7 are physically associated with it to give
supramolecular complex(es) of about 500 kDa as judged from Superdex S200 gel fitration and glycerol gradient ultracentrifugation
experiments. In contrast, the apparent molecular weight of G-CK estimated from an in gel activity assay after SDSPAGE and
renaturation is about 41 kDa. Many of the proteins phosphorylated by and/or associated with G-CK belong to the category of
chaperonines, including HSP90, GRP-94 and −78, and various isoforms of protein disulfide isomerases, suggesting a global role
of this kinase in the modulation of protein folding.
Received 21 October 2005; received after revision 30 November 2005; accepted 6 December 2005
†These authors contributed equally to this work. 相似文献
8.
Gemoll T Habermann JK Lahmann J Szymczak S Lundgren C Bündgen NK Jungbluth T Nordström B Becker S Lomnytska MI Bruch HP Ziegler A Hellman U Auer G Roblick UJ Jörnvall H 《Cellular and molecular life sciences : CMLS》2012,69(2):325-333
DNA aneuploidy has been identified as a prognostic factor in the majority of epithelial malignancies. We aimed at identifying ploidy-associated protein expression in endometrial cancer of different prognostic subgroups. Comparison of gel electrophoresis-based protein expression patterns between normal endometrium (n?=?5), diploid (n?=?7), and aneuploid (n?=?7) endometrial carcinoma detected 121 ploidy-associated protein forms, 42 differentially expressed between normal endometrium and diploid endometrioid carcinomas, 37 between diploid and aneuploid endometrioid carcinomas, and 41 between diploid endometrioid and aneuploid uterine papillary serous cancer. Proteins were identified by mass spectrometry and evaluated by Ingenuity Pathway Analysis. Targets were confirmed by liquid chromatography/mass spectrometry. Mass spectrometry identified 41 distinct polypeptides and pathway analysis resulted in high-ranked networks with vimentin and Nf-κB as central nodes. These results identify ploidy-associated protein expression differences that overrule histopathology-associated expression differences and emphasize particular protein networks in genomic stability of endometrial cancer. 相似文献
9.
10.
目的通过抑郁模型大鼠海马突触的差异蛋白质组学分析,为抑郁症发病机制研究提供依据。方法根据糖水消耗基线值将40只健康雄性Sprague—Dawley大鼠随机分为慢性轻度不可预见性应激组和对照组(每组20只)。通过CUMS实验模式建立大鼠抑郁模型后,运用蔗糖密度梯度离心法提取两组大鼠的海马突触并用透射电镜进行检测。采用传统的双向凝胶电泳和基质辅助激光解析电离飞行时间串联质谱进行差异蛋白质组学分析。结果在行为学评价中CUMS组大鼠糖水消耗量和偏好度均降低(P〈0.01),表明大鼠抑郁模型建立成功。通过结合双向凝胶电泳和质谱分析,共得到6个在CUMS组表达上调和10个在CUMS组表达下调的蛋白质,这些蛋白质的功能主要归为四类,即囊泡调节/递质释放/信号转导、能量代谢、细胞骨架、物质代谢。结论通过对抑郁模型大鼠海马突触的比较蛋白质组学分析及后续的蛋白质功能预测,发现了一些与突触传导功能障碍可能相关的蛋白质,从而为抑郁症发病机制的研究提供了有价值的线索。 相似文献
11.
D. Robinette S. Wada T. Arroll M. G. Levy W. L. Miller E. J. Noga 《Cellular and molecular life sciences : CMLS》1998,54(5):467-475
Three antibacterial proteins were isolated from acid extracts of channel catfish (Ictalurus punctatus) skin by cation exchange chromatography and reverse-phase high-pressure liquid chromatography. The molecular masses of the
proteins were 15.5, 15.5 and 30 kD as determined by SDS-polyacrylamide gel electrophoresis. Mass spectrometry, amino acid
composition and amino acid sequence data suggest that the most abundant protein is closely related to histone H2B. The H2B-like
protein was inhibitory to Aeromonas hydrophila and Saprolegnia spp., which are important bacterial and fungal pathogens of fish. These findings suggest that histones may be important defensive
molecules in fish.
Received 22 December 1997; received after revision 5 March 1998; accepted 5 March 1998 相似文献
12.
目的筛选胎儿宫内发育迟缓大鼠血清中差异表达的蛋白质,寻找与胎儿宫内发育迟缓相关的疾病标志物。方法收集大鼠疾病正常组、IUGR组血浆各3例;TCA法沉淀血清蛋白,建立疾病组和正常组蛋白质的双向凝胶电泳图谱;ImageMaster分析软件分析差异蛋白质;飞行时间质谱(MALDI—TOF—MS)获得差异蛋白肽指纹图谱,MASCOT引擎搜索蛋白质种类。结果鉴定出个差异蛋白点3个,分别为:IGF—I、IGF-Ⅱ、IGFBP-3。结论IGF—I、IGF-II、IGFBP-3,可能是胎儿宫内发育迟缓疾病诊断的生物标志物。 相似文献
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14.
Isolated rat hepatocytes were labeled with 35S-methionine in the presence of 25 mM diethylnitrosamine (DENA). The intrinsically labeled proteins were analyzed by one- and two-dimensional gel electrophoresis and the fluorographic patterns were compared with those obtained from untreated hepatocytes. The results of short term experiments (2 h) show that, in the presence of 25 mM DENA, protein synthesis is inhibited by 50%. This reduction encompasses all protein species without selective inhibition of certain proteins. 相似文献
15.
N Frénoy C Rimbant D Buffe 《Comptes rendus des séances de l'Académie des sciences. Série D, Sciences naturelles》1975,281(21):1649-1651
alpha2 H globulin, a glycoferroprotein, was first demonstrated in the sera of patients with malignant diseases. This protein was isolated from cancerous human liver, and compared with ferritin, a ferroprotein showing some identical properties (presence of iron, high molecular weight, common antigenic determinants). However, physicochemical differences were observed between these two proteins. The study of protein dissociation was performed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate after reduction by mercaptoethanol. A similar molecular weight of 19 000 is obtained for subunits of these two proteins. This value agrees well with the results obtained by other authors for ferritin. 相似文献
16.
Zhang LJ Wang XE Peng X Wei YJ Cao R Liu Z Xiong JX Yin XF Ping C Liang S 《Cellular and molecular life sciences : CMLS》2006,63(15):1790-1804
To characterize low-copy integral membrane proteins and offer some methods for human liver proteome projects, we fractionated
highly purified rat liver plasma membrane (PM). PM was purified through two sucrose density gradient centrifugations, and
treated with 0.1 M Na2CO3, chloroform/methanol and Triton X-100. Proteins were separated by electrophoresis and submitted to mass spectrometry analysis.
Four hundred and fiftyseven non-redundant membrane proteins were identified, of which 23% (105) were integral membrane proteins
with one or more transmembrane domains. One hundred and fifty-three (33.5%) had no location annotation and 68 were unknown-function
proteins. The proteins from different fractions were complementory. A database search for all identified proteins revealed
that 53 proteins were involved in the cell communication pathway. More interestingly, more than 50% of the proteins had a
protein abundance index concentration of less than 0.1 mol/l, and 12% proteins a concentration 100 times less than that of
arginase 1 and actin.
Received 15 March 2006; received after revision 17 May 2006; accepted 10 June 2006
L.-J. Zhang and X.-e Wang are contributed equally to this work. 相似文献
17.
Biochemical characterization and bacterial expression of an odorant-binding protein from <Emphasis Type="Italic">Locusta migratoria</Emphasis> 总被引:1,自引:0,他引:1
Ban L Scaloni A D'Ambrosio C Zhang L Yahn Y Pelosi P 《Cellular and molecular life sciences : CMLS》2003,60(2):390-400
Analysis of soluble proteins from different body parts of Locusta migratoria revealed a fast-migrating component in native electrophoresis, unique to antennae of both sexes. N-terminal sequence analysis and cloning identified this protein as a member of the insect odorant-binding proteins, carrying a well-conserved six-cysteine motif. Mass spectrometry analysis confirmed the occurrence of two distinct polypeptide species determined by nucleotide sequencing and demonstrated that the cysteine residues are paired in an interlocked fashion. The protein was expressed in a bacterial system with yields of about 10 mg/l of culture, mostly present as inclusion bodies. However, this recombinant product was solubilized after disulfide reduction. Air oxidation yielded a species with all disulfides spontaneously formed as in the native counterpart. Both native and recombinant proteins migrated as a dimer in gel filtration chromatography. Ligand binding was measured, using N-phenyl-1-naphthylamine as the fluorescent probe; the affinity of other ligands was measured in competitive binding assays. The protein exhibited great resistance to thermal denaturation even following prolonged treatment at 100 degrees C. A structural model for this dimeric species was generated on the basis of its sequence homology with Bombyx mori pheromone-binding protein, whose three-dimensional structure has been resolved as an unbound species and in complex with its physiological ligand. This is the first report of an odorant-binding protein identified and characterized from Orthoptera. 相似文献
18.
Gemoll T Roblick UJ Szymczak S Braunschweig T Becker S Igl BW Bruch HP Ziegler A Hellman U Difilippantonio MJ Ried T Jörnvall H Auer G Habermann JK 《Cellular and molecular life sciences : CMLS》2011,68(19):3261-3274
DNA aneuploidy has been identified as a prognostic factor for epithelial malignancies. Further understanding of the translation of DNA aneuploidy into protein expression will help to define novel biomarkers to improve therapies and prognosis. DNA ploidy was assessed by image cytometry. Comparison of gel-electrophoresis-based protein expression patterns of three diploid and four aneuploid colorectal cancer cell lines detected 64 ploidy-associated proteins. Proteins were identified by mass spectrometry and subjected to Ingenuity Pathway Analysis resulting in two overlapping high-ranked networks maintaining Cellular Assembly and Organization, Cell Cycle, and Cellular Growth and Proliferation. CAPZA1, TXNL1, and HDAC2 were significantly validated by Western blotting in cell lines and the latter two showed expression differences also in clinical samples using a tissue microarray of normal mucosa (n?=?19), diploid (n?=?31), and aneuploid (n?=?47) carcinomas. The results suggest that distinct protein expression patterns, affecting TXNL1 and HDAC2, distinguish aneuploid with poor prognosis from diploid colorectal cancers. 相似文献
19.
J. van der Velden P. Borgdorff G. J. M. Stienen 《Cellular and molecular life sciences : CMLS》1999,55(5):788-798
To investigate whether during cardiac hypertrophy changes occur in contractile protein composition and in mechanical and
energetic properties of the myocardium, contractile protein composition, isometric force and adenosine triphosphate (ATP)
consumption were studied in control and hypertrophied guinea-pig hearts. Cardiac hypertrophy was induced by adding minoxidil
(120 or 200 mg/l) to the drinking water. Protein analysis was performed by one-dimensional gel electrophoresis. The myosin
heavy-chain (MHC) composition was determined in an enzyme-linked immunosorbent assay (ELISA). ATP consumption and force development
were simultaneously measured during isometric contraction in chemically skinned trabeculae. Histochemical analysis of cross-sectional
area of cardiomyocytes and interstitial space was performed on the left ventricular tissue of 200 mg/l minoxidil-treated and
control guinea pigs. Minoxidil treatment (120 and 200 mg/l) significantly increased left ventricular dry weight normalized
for body weight by 19 ± 4 and 24 ± 4%, respectively. No significant differences were found in the cellular cross-sectional
area, while interstitial space was slightly decreased in minoxidil-treated hearts. In left ventricular trabeculae of 200 mg/l
minoxidil-treated guinea pigs, ATPase activity was slightly less than in those of control guinea pigs, whereas force did not
differ significantly. Calcium sensitivity of force and ATPase activity were not affected by minoxidil treatment. Gel electrophoresis
revealed no difference in contractile protein composition, but a tendency towards a lower amount of α-MHC in the minoxidil-treated
hearts was found in ELISA.
Received 1 February 1999; accepted 15 March 1999 相似文献