This study was performed to comprehend the plasma proteins expressed specifically during early pregnancy in pregnant or non-pregnant Hanwoo using proteomic analysis technique. Plasma samples (0, 2, 3, 4, 7, and 11 weeks after AI) were obtained from pregnant (P, n=3) or non-pregnant (NP, n=4) Hanwoo, respectively. To evaluate proteins differentially expressed, 2-dimensional electrophoresis (2DE) was conducted. Normalized protein spots were selected for the significant expression variation deviated over two fold in its expression level between two groups. Molecular functions of the proteins were DNA binding, protein binding, hemoglobin binding, ferrochelatase and transporter activity and arylestera, respectively. According to western blotting, haptoglobin was specifically expressed only in NP group during early pregnancy; however, paraoxonase 1 was highly expressed in pregnant group. Based on these results, pregnancy was maintained successfully by the activation of specific plasma proteins associated with immune system and antioxidant regulation during early pregnancy in Hanwoo

Pluripotency of human embryonic stem cell (hESC) is one of the most valuable ability of hESCs for applying cell therapy field, but also showing side effect, for example teratoma formation. When transplant multipotent stem cell, such as mesnchymal stem cell (MSC) which retains similar differentiation ability, they do not form teratoma in vivo, but there exist limitation of cellular source supply. Accordingly, differentiation of hESC into MSC will be promising cellular source with strong points of both hESC and MSC line. In this study, we described the derivation of MSC like cell population from feeder free cultured hESC (hESC- MSC) using direct differentiation system. Cells population, hESC-MSC and bone marrow derived MSC (BM-MSC) retained similar characteristics in vitro, such as morphology, MSC specific marker expression and differentiation capacity. At the point of differentiation of both cell populations, differentiation rate was slower in hESC-MSC than BM-MSC. As these reason, to verify differentially expressed molecular condition of both cell population which bring out different differentiation rate, we compare the molecular condition of hESC-MSC and BM-MSC using 2-D proteomic analysis tool. In the proteomic analysis, we identified 49 differentially expressed proteins in hESC-MSC and BM-MSC, and they involved in different biological process such as positive regulation of molecular function, biological process, cellular metabolic process, nitrogen compound metabolic process, macromolecule metabolic process, metabolic process, molecular function, and positive regulation of molecular function and regulation of ubiquitin protein ligase activity during mitotic cell cycle, cellular response to stress, and RNA localization. As the related function of differentially expressed proteins, we sought to these proteins were key regulators which contribute to their differentiation rate, developmental process and cell proliferation. Our results suggest that the expressions of these proteins between the hESC-MSC and BM-MSC, could give to us further evidence for hESC differentiation into the mesenchymal stem cell is associated with a differentiation factor. As the initial step to understand fundamental difference of hESC-MSC and BM-MSC, we sought to investigate different protein expression profile. And the grafting of hESC differentiation into MSC and their comparative proteomic analysis will be positively contribute to cell therapy without cellular source limitation, also with exact background of their molecular condition.

tasks that require nematode extraction and microscopic examination. To develop a more efficient detection method for Bursaphelenchus xylophilus, we first generated monoclonal antibodies (MAbs) specific to B. xylophilus. Among 2,304 hybridoma fusions screened, a hybridoma clone named 3-2A7-2H5 recognized a single protein from B. xylophilus specifically. We finally selected the MAb clone 3-2A7-2H5-D9-F10 (D9-F10) for further studies. To identify the antigenic target of MAb-D9-F10, we analyzed proteins in spots, fractions or bands via nano liquid chromatography electrospray ionization quadrupole ion trap mass spectrometry (nano-LC-ESI-Q-IT-MS). Peptides of galactose-binding lectin-1 of B. xylophilus (Bx-LEC-1) were commonly detected in several proteomic analyses, demonstrating that this LEC-1 is the antigenic target of MAb-D9-F10. The localization of MAb-D9-F10 immunoreactivities at the area of the median bulb and esophageal glands suggested that the Bx-LEC-1 may be involved in food perception and digestion. The Bx-LEC-1 has two non-identical galactose-binding lectin domains important for carbohydrate binding. The affinity of the Bx-LEC-1 to D-(+)-raffinose and N-acetyllactosamine were much higher than that to L-(+)-rhamnose. Based on this combination of evidences, MAb-D9-F10 is the first identified molecular biomarker specific to the Bx-LEC-1.

There are increasing interests in developing methods specifically detecting pathogenic Bursaphelenchus xylophilus. In order to develop a detecting method for B .xylophilus, at first we generated monoclonal antibodies (MAbs) specific to B. xylophilus, discriminating from other pine tree resident nematodes. Among 2304 hybridoma fusions screened. We finally selected a MAb clone, 9F10 and used for further study. To identify the antigenic target of MAb-9F10, we employed several biochemical methods such as SDS-PAGE, 2 dimensional electrophoresis, anion exchange chromatography, and immunoprecipitation to separate and isolate an antigenic target. Proteins from above methods were analyzed via nano-LC-ESI-Q-IT-MS. Peptides of GaLECtin were always detected from several proteomic analyses, suggesting that GaLECtin is the antigenic target of MAb-9F10.

Upon oviposition, parasitoid wasps inject their eggs along with venom, teratocytes and polydnavirus (PDV) on the host. Among these parasitic factors, PDVs are known to suppress the host immune system and utilize the host translational mechanisms allowing the juvenile parasitoid to develop. Polydnavirusencoded genes can selectively inhibit host translation and still use the translation machinery of the host to synthesize their own proteins. In this study, we utilize a proteomic approach involving two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) that couples isoelectric focusing (IEF) and SDS-PAGE to resolve complex protein mixtures that results from the parastization of Cotesia plutellae on the lepidopteran host, Plutella xylostella. We specifically analyze the changes in protein synthesis using this technique after treatment of HTIFs that has been previously identified on C. plutellae. The difference in protein profile due to parasitization was confirmed by in vitro translation assay using rabbit reticulosyte lysate.

Cyclosporin A (CsA) has been used clinically as an immunosuppressive drug to prevent organ transplant rejection and in basic research as a mitochondrial permeability blocker. It has been reported that CsA has a protective role in severed neurons and a neurotrophic effect in neuronal cells. However, the molecular mechanisms underlying the stimulation of neuronal cell proliferation by CsA have not yet been elucidated. In our current study, we investigated CsA responsive proteins in PC12 cells using a systematic proteomic approach. The viability of these cells following CsA treatment increased in a dose- and time-dependent manner. Proteins in the CsA-treated PC12 cells were profiled by two-dimensional gel electrophoresis (2-DE) and identified by matrix-assisted laser desorption ionization time-of flight (MALDI-TOF) and electrospray ionization quadupole time-of-flight mass spectrometries (EIQ-TOFMS). This differential expression analysis showed significant changes for 10 proteins (6 up-regulated and 4 down-regulated) upon CsA treatment that were related to cell proliferation, metabolism and the stress response. These proteomics data further our understanding of the proliferation mechanisms of PC12 cells exposed to CsA and demonstrate that our methodology has potential to further elucidate the mechanisms and pathways involved.

This study was performed to identify and analyze the specifically expressed plasma proteins during early pregnancy in both pregnant and non-pregnant Hanwoo. Blood samples were collected at 0 (the day of AI), 2, 3, 4, 7, and 11 weeks after AI from pregnant (n=3) and non-pregnant (n=4) Hanwoo, respectively. The hematological parameters were measured. After 2-dimensional electrophoresis using serum, normalized protein spots were selected for the significant expression variation deviated over two fold in its expression level between two groups. Among 17 spots selected, 15 were identified as albumin, IgG1 heavy chain constant region, haptoglobin, ferrochelatase, fibrinogen, hemopexin. 5 spots were expressed only in non-pregnant specific. The spot identification of 1105 and 6106 was decreased after 3 weeks from AI. However, 2/17 spots were still unidentified. Further studies are needed to analyze the function of the proteins associated with early pregnancy.

Human saliva contains a large number of proteins and peptides whose composition may alter as a consequence 。f disease. To date. however‘ the proteins and peptides that routinely populate thi s ora l fluid a re largely unknown To provide a catalogue of saliva proteins, we have surveyed the unstimulated human whole saliva by us ing shotgun proteornics. For the shotgun approach, whole saliva proteins were digested into peptides with ChemDigestD ‘ and the resul ting peptide fragments were separated by RP-HPLC, followed by each fraction was t ryptic digestion. ChemDigestD-Trypsin digested peptides were analyzed by tandem mass spectrometry(MS/MS) us ing a nano-LC eq 버 pped quadrupole-time of f1ight mass spectrometer, and the obtained spectra were searched against human protein seq uence data base using MASCOT. Shotgun proteomics allowed a total of 291 human pr。 teins to be confidently assigned . The largest group(17 .2%) of the identified proteins sorted into functional catego ries was included in t he signal t ransduction funct ion except for the hypothetical 0 1' unknown function. This work provides a valuable s ta rting point for the analysis of human salivary proteins and their biological functions and candidates from human whole sali va that may prove to be of diagnostic and therapeutic significance

The human ELAV(embryonic lethal abnormal vision)-like protein HuR stabilizes a certain group of cellular mRNAs that contain AU- ri ch elements in theil‘ 3’• untranslated region, Dysregulation of mRNA s tability may be relevant in tumor biology and may lead to abnormal expression of several proteins in malignant tumors, The aim of this study is to identify the differentially expressed proteins according to the functi onal activi ty of HuR Methods : We used stabl e expression of small interfering RNA(siRNA) of HuR gene to inhibit the expression of HuR in human ora l car cinoma cell lines‘ KB cell line and YD10B cell line‘ and compared the proteomic changes between s iRNA-treated and cont rol cell line using two-dimensional gel electrophoresis , Flow cytometry caliber scan(FACS) was employed to investigate the effects of HuR on cell apoptosis and proliferation , Results: Seventeen differentially expressed proteins between the two cell lines were identified by electrospray ioruzati on quadrupole time-of- fl ight mass spectrometry(ESI-Q-TOF MS) and database searching, Among them there are eleven proteins which are significant ly up- regulated in siRNA- treated cell line, which include heat shock protein 10(influencing nucl eocytoplas rnic transpo 다， cell dedifferentiation , and inhibition of apoptosis) , keratin 19(basal cell differ ent iat ion) ‘ and nucleoside diphosphate kinase B(G protein activator), etc Enolase 1 and Ml of pyruvate kinase a re the representatives of signifi cantly down-regulated proteins in siRNA-treated cell 11l1e Conclusion : Our data suggest that HuR participate in mRNAs stability of proteins that have the counter effects in the carcinogenesis of oral ca ncer , And the functiona l proteomics a re needed to elucidate the detailed interactions between HuR and t hese molec ules

Human saliva conta ins a la rge number of proteins and peptides whose composition may alter as a conseq uence of disease. '1'0 date‘ however. the proteins and peptides that routinely populate t his oral fluid are largely unknown, '1'0 provid e a ca ta logue 이， sali va protei ns. we have surveyed the unstimulated human whole saliva by using shotgun proteomics. F'or the shotgun a pproach‘ whole sali va proteins were digested into peptides with ChemDigestD and the res ulting pe ptide fragments were sepa rated by RP- HPLC, followed by each fraction was tryptic di gestion ChemDiges tD- Trypsin diges ted pe pt ides were analyzed by tandem mass spectrometry (MS!MS) using a nano-LC equi pped quadru po le-time of fli ght rnass spect rometer, and the obtained spectra were searched against human protein sequence da tabase us ing MASCOT Shotgun proteomics a llowed a total of 291 human proteins to be confidently assigned. The largest gro u p (17 , 2%) of the identifi ed proteins sorted into functional categories was included in the s ignal transducti on function except for the hypothet ical or unknown functio n, This work provides a valuable starting point for the ana lys is of human sa l i va ry protei ns a nd theil‘ biological functions and candidates from human whole saliva that may prove to be of diagn ost ic and t herapeutic s ignif‘ Ica nce