Analysis of lysosomal membrane proteins exposed to melanin in HeLa cells

OBJECTIVES: There have been developed to use targeting ability for antimicrobial, anticancerous, gene therapy and cosmetics through analysis of various membrane proteins isolated from cell organelles. METHODS: It was examined about the lysosomal membrane protein extracted from lysosome isolated from HeLa cell treated by 100 ppm melanin for 24 hours in order to find associated with targeting ability to melanin using by 2-dimensional electrophoresis. RESULTS: The result showed 14 up-regulated (1.5-fold) and 13 down-regulated (2.0-fold) spots in relation to melanin exposure. CONCLUSIONS: It has been found that lysosomal membrane proteins are associated with melanin to decolorize and quantity through cellular activation of lysosome.

Proteomic Analysis of a Rat Cerebral Ischemic Injury Model after Human Cerebral Endothelial Cell Transplantation

OBJECTIVE: Cerebral endothelial cells have unique biological features and are fascinating candidate cells for stroke therapy. METHODS: In order to understand the molecular mechanisms of human cerebral endothelial cell (hCMEC/D3) transplantation in a rat stroke model, we performed proteomic analysis using 2-dimensional electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Protein expression was confirmed by quantitative real-time PCR and Western blot. RESULTS: Several protein spots were identified by gel electrophoresis in the sham, cerebral ischemia (CI), and CI with hCMEC/D3 treatment cerebral ischemia with cell transplantation (CT) groups, and we identified 14 differentially expressed proteins in the CT group. Proteins involved in mitochondrial dysfunction (paraplegin matrix AAA peptidase subunit, SPG7), neuroinflammation (peroxiredoxin 6, PRDX6), and neuronal death (zinc finger protein 90, ZFP90) were markedly reduced in the CT group compared with the CI group. The expression of chloride intracellular channel 4 proteins involved in post-ischemic vasculogenesis was significantly decreased in the CI group but comparable to sham in the CT group. CONCLUSION: These results contribute to our understanding of the early phase processes that follow cerebral endothelial cell treatment in CI. Moreover, some of the identified proteins may present promising new targets for stroke therapy.

Initial research of screening for the differentially expressed proteins in beagles irradiated with 4.5 Gy60 CO γ-rays by two-dimensional gel electrophoresis and mass spectrometry / 中华放射医学与防护杂志

Objective To explore the mechanisms of cytokines on acute radiation disease in irradiated beagles.Methods The sera of beagles irradiated with 4.5 Gy γ-rays with cytokines treatment was collected at different time points post irradiation.The two-dimensional gel electrophoresis(2-DE)was used to isolate and compare the differentially expressed proteins in sera.HD-MS was used to analyze the differentially expressed proteins with significance,and the amino acid sequences should be determined. Results High resolution 2-DE gel map was obtained.There were six differentially expressed proteins in sera of irradiated beagles at different time points.Four protein spots were successfully identified by MS.A significant spot was identified as serum amyloid A(SAA)by HD-MS,with relative molecular mass of 13 077 and isoelectfie point of 6.26.Expression of SAA was not found 1 d pre-irradiation and 36 d postirradiation,but increased slightly 1 d(0.2166)and significantly 14 d post-irradiation(0.4577). Conclusions The expression of serum amyloid A was consistent with the process of acute radiation injury,which might indicate the turnover of the disease.

Proteomic Approach in Gastrointestinal and Liver Research / 대한소화기학회지

In the post-genomic era, the focus of research is now moving to functional genomics employing the information on predicted gene products provided by genome sequencing. Proteomics, the global analysis of structures, functions, and interactions of whole cellular proteins, draws the special attention as a tool for documenting the disease pathogenesis or progression. The high-throughput technology has become feasible by considerable improvement of two dimensional electrophoresis and mass fingerprinting. Thus proteome techniques can be used as tools to study the disease processes, develop new biomakers for diagnosis and early detection of diseases, and accelerate drug development. In this review, we discuss the background and techniques of proteomics, and potential applications to the research of gastrointestinal diseases.

Application of Proteomics in the Study of Mycobacterium tuberculosis / 微生物学通报

Proteomics is an emerging discipline developed on the basis of genomics.The fundamental techniques of proteomics include sample preparation,protein separation,protein identification and analysis,and its core techniques are two-dimensional gel electrophoresis and mass spectrometry.In recent years,proteomics has been used in researching the field of Mycobacterium tuberculosis(MTB).Proteomics promotes deep understanding of the pathogenesis of MTB and resistance mechanism via isolating,identifying and analyzing the whole-cell protein and secreted proteins.The development of new vaccine against MTB has showed some promising results based on proteomics.Some powerful early diagnostic markers have been discovered via analyzing the protein composition of MTB clinical isolates.Proteomics also applies to find potential new drug targets,and it has shown many valuable research productions in developing new an-ti-MTB drugs.In summary,the application of proteomics has built a solid foundation for the development of prevention,early diagnosis and treatment of tuberculosis.