1.Improved Algorithms for the Identification of Yeast Proteins and Significant Transcription Factor and Motif Analysis.
Seung Won LEE ; Seong Eui HONG ; Kyoo Yeol LEE ; Do Il CHOI ; Hae Young CHUNG ; Cheol Goo HUR
Genomics & Informatics 2006;4(2):87-93
With the rapid development of MS technologiesy, the demands for a more sophisticated MS interpretation algorithm haves grown as well. We have developed a new protein fingerprinting method using a binomial distribution, (fBIND). With the fBIND, we improved the performance accuracy of protein fingerprinting up to the maximum 49% (more than MOWSE) and 2% than(at a previous binomial distribution approach studied by of Wool et al.) as compared to the established algorithms. Moreover, we also suggest a the statistical approach to define the significance of transcription factors and motifs in the identified proteins based on the Gene Ontology (GO).
Binomial Distribution
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Fungal Proteins*
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Gene Ontology
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Peptide Mapping
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Transcription Factors*
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Wool
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Yeasts*
2.Comparison of proteomics between acute myeloid leukemia and acute lymphoid leukemia.
Ping XIAO ; Yao-Ying ZENG ; Yan-Fang NIE ; Wei LIN
Journal of Experimental Hematology 2011;19(6):1353-1356
The aim of this study was to explore the distinct protein profiles of different subtype of acute myeloid leukemia (AML), including M(1), M(2), M(3) and acute lymphoid leukemia (ALL) by differential proteomic expression analysis. The proteins of bone marrow leukemia cells from AML and ALL patients were separated by two-dimensional electrophoresis (2-DE). 2-DE patterns were analyzed by PDQuest 7.4 software and the differentially expressed proteins were identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) and bioinformatics. The results indicated that 21 differentially expressed proteins were found by 2-DE and 15 were identified by MS to be significantly differentially expressed. In AML, seven proteins were highly expressed such as MPO, PRDX3, CALR and ECH1 and so on, and eight proteins were highly expressed in ALL, including ARHGDIB, PFN1 and ACTG1 and so on. It is concluded that the distinct protein profiles between AML and ALL have been proved. It may be helpful for the identification of new targets for specific treatment approaches and the molecular markers for the early diagnosis of leukemia.
Humans
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Leukemia, Myeloid, Acute
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metabolism
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Peptide Mapping
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Precursor Cell Lymphoblastic Leukemia-Lymphoma
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metabolism
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Proteome
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Proteomics
3.Commentary on: Mobley AJ, Lam YW, Lau KM, Pais VM, Lesperance JO, Steadman B, et al. Monitoring the serological Proteome Colon, the latest modality in prostate cancer detection. J Urol 2004; 172: 331-7.
Asian Journal of Andrology 2004;6(4):283-283
Biomarkers
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Humans
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Male
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Peptide Mapping
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Prognosis
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Prostatic Neoplasms
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blood
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Proteome
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metabolism
4.Evaluating peptide mass fingerprinting-based protein identification.
Senthilkumar DAMODARAN ; Troy D WOOD ; Priyadharsini NAGARAJAN ; Richard A RABIN
Genomics, Proteomics & Bioinformatics 2007;5(3-4):152-157
Identification of proteins by mass spectrometry (MS) is an essential step in proteomic studies and is typically accomplished by either peptide mass fingerprinting (PMF) or amino acid sequencing of the peptide. Although sequence information from MS/MS analysis can be used to validate PMF-based protein identification, it may not be practical when analyzing a large number of proteins and when high- throughput MS/MS instrumentation is not readily available. At present, a vast majority of proteomic studies employ PMF. However, there are huge disparities in criteria used to identify proteins using PMF. Therefore, to reduce incorrect protein identification using PMF, and also to increase confidence in PMF-based protein identification without accompanying MS/MS analysis, definitive guiding principles are essential. To this end, we propose a value-based scoring system that provides guidance on evaluating when PMF-based protein identification can be deemed sufficient without accompanying amino acid sequence data from MS/MS analysis.
Algorithms
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Peptide Mapping
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methods
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statistics & numerical data
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Proteomics
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methods
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statistics & numerical data
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Tandem Mass Spectrometry
5.Epitope screening of influenza A (H3N2) by using phage display library.
Yan-Wei ZHONG ; Dong-Ping XU ; Xiao-Dong LI ; Jiw-Zeng DAI ; Biao XU ; Le LI
Chinese Journal of Experimental and Clinical Virology 2009;23(4):272-274
OBJECTIVETo screen the influenza A (H3N2) mimotopes by using phage display library.
METHODSUsing influenza A (H3N2) monoclonal antibody as selective molecule, a 7 mer phage peptide library was biopanned and positive clones were selected by ELISA, competition assay and DNA sequencing.
RESULTS21 positive clones were chosen for DNA sequencing. From the experiment and sequencing comparison results, one epitope was comfirmed as mimotope of influenza A (H3N2).
CONCLUSIONInfluenza A (H3N2) mimotope was obtained by phage peptide library screening. The result provide a new approach for new Influenza virals vaccine development.
Epitope Mapping ; Humans ; Influenza A Virus, H3N2 Subtype ; chemistry ; genetics ; immunology ; Peptide Library
6.Establishment of 2-dimensional gel electrophoresis map and analysis of proteomics from human nasal polyps.
Guang-xiang HE ; Hong SUN ; Tian-sheng WANG ; Gui LI ; Huo-wang LIU ; Yu CHEN
Journal of Central South University(Medical Sciences) 2006;31(4):487-492
OBJECTIVE:
To establish 2-dimensional polyacrylamide gel electrophoresis (2-DE) map from human nasal polyps and normal nasal mucosa, and to identify differential expression proteins of 2-DE map.
METHODS:
Samples of nasal polyps and nasal mucosa (each sample group containing 7 cases) were obtained. The total proteins were extracted and separated by immobilized pH gradient (IPG)-based 2-DE. The silver-stained 2-DE was scanned with digital Imagescanner and analyzed with ImageMaster 2-DE Elite 4.01 software. To obtain peptide mass fingerprint (PMF) of differential protein spots, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was used. The PMF was searched in Swiss-Prot and TreMBL database by Pept-Ident software, to identify differential expression proteins.
RESULTS:
The well-resolved, reproducible 2-DE maps of nasal polyps and nasal mucosa were established. For the polyps tissues, the average proteins spot of three 2-DE maps was 825+/-78; and 682+/-96 spot was matched with the average matching rate of 82.7%. The average deviations of matched spot position were (1.13+/-0.16) mm in IEF direction and (1.45+/-0.21) mm in SDS-PAGE direction, respectively. For the nasal mucosa tissues, the average proteins spot of three 2-DE maps was 936+/-62; and 821+/-78 spots were matched with the average matching rate of 87.7%. After comparing the 2-DE maps of nasal polyps and nasal mucosa tissues, the protein spots were 1,458 and 1,617 respectively; and 1,026 protein spots were matched. Forty differential expression protein spots were incised from silver staining gel randomly and digested in the gel by TPCK-Trypsin. Thirty-four PMFs were obtained by MALDI-TOF-MS and 24 differential proteins were identified.
CONCLUSION
The well-resolved, reproducible 2-DE maps of human nasal polyps and nasal mucosa have been successfully established. Certain differential proteins related to the pathogenesis of human nasal polyps are identified.
Adult
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Electrophoresis, Gel, Two-Dimensional
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Female
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Humans
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Male
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Middle Aged
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Nasal Polyps
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metabolism
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Peptide Mapping
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Proteomics
7.Two-dimensional polyacrylamide gel electrophoresis-based serum protein separation: comparison of two sample preparation methods.
Liang ZHAO ; Yan-Qing DING ; Li LIANG ; Xin LI ; Xue-Hua LI ; Li-Sha WU
Journal of Southern Medical University 2007;27(1):5-8
OBJECTIVETo evaluate the effects of two sample preparation methods on two-dimensional polyacrylamide gel electrophoresis (2-DE)-based serum protein separation, and produce high-resolution and reproducible 2-DE images for identifying disease-related serum protein.
METHODSDirect solubilization and hot SDS methods were used separately to extract and handle the total proteins of serum samples from patients with colorectal carcinoma. Immobilized pH gradient 2-DE was used to separate the total proteins. After image analysis of silver-stained 2-D gels, 3 differential protein spots were identified by matrix-assisted laser desorption/time-of-flight mass spectrometry.
RESULTSThe total proteins treated with hot SDS method were used to perform 2-DE. 2-DE patterns with high resolution and reproducibility were obtained for human serum samples. 2-DE was performed 3 times for the samples treated by direct solubilization and hot SDS methods, respectively, resulting in the average number of spots of 675-/+46 and 702-/+49, respectively. The average matching protein spots were 573-/+42 and 623-/+52, with average matching rate of 85.3% and 89.6%, respectively. The average position deviation of matched spots in different gels was 0.85-/+0.30 mm and 0.81-/+0.28 mm in IEF direction, and 1.02-/+0.18 mm and 0.97-/+0.12 mm in SDS-PAGE direction. Mass spectrometry of the 2-D gels treated with hot SDS method generated high-quality mass spectra, and the sample preparation method allowed detection of relatively low abundance protein.
CONCLUSIONHot SDS method is more effective for human serum protein sample preparation and well-resolved, reproducible 2-DE profiles of human serum have been established in this study.
Blood Protein Electrophoresis ; Blood Proteins ; analysis ; isolation & purification ; Electrophoresis, Gel, Two-Dimensional ; methods ; Electrophoresis, Polyacrylamide Gel ; methods ; Humans ; Peptide Mapping ; Protein Interaction Mapping ; Reproducibility of Results
8.Proteomic Analysis of Helicobacter pylori Whole Cell Proteins using the Narrow Range IPG Strips.
Jeong Won PARK ; Seung Gyu LEE ; Jae Young SONG ; Jin Su JUN ; Jung Soo JOO ; Hee Shang YOUN ; Ji Hyun SEO ; Hyung Lyun KANG ; Seung Chul BAIK ; Woo Kon LEE ; Myung Je CHO ; Kwang Ho RHEE
Journal of Bacteriology and Virology 2007;37(4):203-212
It has been reported that most of Helicobacter pylori proteome components appear so crowded in the region of pH 4.5~8.0 that a lot of them were inseparable in 2-DE using the broad range IPG strip. Therefore, inseparable protein spots in 2-DE profiles have to be apart from each other for improving the protein identification. Here, we attempt to examine the usability of the narrow range IPG strips for separating close spots in the broad range IPG strip at proteomic analysis of H. pylori. The whole cell proteins of H. pylori strain 26695 were separated by narrow range IPG strips (pI 3.9~5.1, 4.7~5.9, 5.5~6.7, and 6.3~8.3, respectively), followed by SDS-PAGE, and visualized by silver staining, showing that the distances between spots were widened and the total number of detectable spots was increased. Resolved protein spots were identified by the peptide fingerprinting using MALDI-TOF-MS. As a result, 87 expressed proteins were identified by the peptide fingerprinting. Of them, 23 proteins, including hydrogenase expression/formation protein, purine-binding chemotaxis protein, and ribosomal protein S6, have not been reported in the previous proteome studies of H. pylori. Thus, these results demonstrate that the high complexity proteome components could be effectively separated using the narrow range IPG strips, which might be helpful to strengthen the contents of the master protein map of the H. pylori reference strain.
Chemotaxis
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Electrophoresis, Polyacrylamide Gel
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Helicobacter pylori*
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Helicobacter*
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Hydrogen-Ion Concentration
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Hydrogenase
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Peptide Mapping
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Proteome
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Proteomics
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Ribosomal Protein S6
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Silver Staining
9.In vitro display technologies.
Song YAN ; Yi ZHANG ; Hongli LU ; Xuewei DONG ; Chao TANG ; Jun MU
Journal of Biomedical Engineering 2009;26(6):1367-1371
The application of in vitro selection method to isolate nucleic acids, peptides and proteins according to their functions has been studied intensively in recent years. In vitro display technologies are not limited by cellular transformation efficiencies; thus, very large libraries of up to 10(13)-10(14) members can be built. The most popular in vitro display technologies are ribosome display and mRNA display; ribosome display achieves the mRNA-ribosome-nascent peptide complexes by stalling the translating ribosome in an in vitro translation reaction. In mRNA display, the mRNA-protein complex is achieved by binding the two macromolecules through a small adaptor molecule, typically puromycin; these mRNA-peptide fusions can then be purified and subjected to in vitro selection. In vitro display technologies provide a different approach to the in vitro selection and directed evolution of peptides and proteins. This review focuses on the principle and method of ribosome display and mRNA display technologies, and discusses their applications.
Animals
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Directed Molecular Evolution
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Gene Library
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Humans
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Peptide Library
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Protein Interaction Mapping
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methods
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RNA, Messenger
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chemistry
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genetics
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Ribosomes
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chemistry
;
genetics
10.Application of proteomics in the research of discrepancy proteins in gastric cancer.
Bo MA ; Ji-sheng ZHAO ; Da-yong DING
Chinese Journal of Gastrointestinal Surgery 2006;9(6):534-537
OBJECTIVETo explore the discrepancy proteins in gastric cancer by proteome analysis.
METHODSTotal proteins of gastric cancer tissues and matched normal gastric epithelial tissues were separated respectively by two-dimensional polyacrylamide gel electrophoresis (2-DE). Mass spectrometry was used to test the differentially expressed proteins.
RESULTSOne thousand one hundred and forty-seven protein spots from gastric cancer tissue and 1079 spots from the normal tissue were gained. Out of 164 different protein spots, 41 were only expressed in gastric cancer tissue, 27 were unique in normal tissue, 39 were up-regulated and 57 were down-regulated in gastric cancer. Seven proteins, which were highly expressed in gastric cancer tissue, were identified.
CONCLUSIONDifferent protein spots between gastric cancer tissues and normal gastric epithelial tissue were gained by proteomics. The 7 discrepancy proteins were further identified. It establishes the foundation of finding specific gastric cancer proteins, which act as biomarkers for the diagnosis and prognosis of gastric cancer.
Case-Control Studies ; Databases, Protein ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Mass Spectrometry ; Peptide Mapping ; Proteomics ; methods ; Stomach Neoplasms ; metabolism