Two-dimensional electrophoresis reveals differential protein expression in high-and low-secreting variants of the rat basophilic leukaemia cell lineThe identification of cellular proteins: - High secretory phenotype on subclones of the rat basophilic leukaemia(RBL) cell line - 2DE, silver staining - Coomassie blue staining, DE-MALDI-MS - GTP overlay and immunoblotting : Differential expression of small GTP binding- proteins, including Rab3 proteins, in high- and low-secreting clones1. Introduction* Mast cell lineage and basophils in allergic reaction: Receptors of IgE , cross-linking of FcεRl, Including histamine, proteoglycans and endoproteases Differences in secretory phenotype: Change in the expression of proteins * The establishment of a link between the differential expression of specific proteins and altered secretory phenotype: Rational therapeutic interventions inhibiting mast cell/basophil degranulation 2-DE, DE-MALDI-MS, GTP overlay, immunoblotting2.Materials and methods1. Reagents 2. Cell culture : RBL-2H3.1, RBL-2H3.2 cells 3. Sample preparation 4. 2-DE 5. Silver staining of second-dimensional gels 6. Coomassie blue staining 7. DE-MALDI-MS 8.〔α-32P〕GTP overlay of 2-D gels 9. Immunochemical detection of Rab3 protein3.Results and discussion1. 2-D electrophoretic profiles and DE- MALDI 2.〔α-32P〕GTP overlay of 2-D gelsFig1.RBL-2H3.1 cells: 2-DE pattern obtained after silver staining2-DE gel pattern after Coomassie blue stainingFig2.RBL-2H3.2 cells:2-DE pattern obtained after silver staining2-DE gel apttern after Coomassie blue stainingBoxed region of the silver-stained gel shown in Fig 1, Fig 2GTP-overlayImmunoblotting with anti-Rab3 antibody4. Concluding remarks* 2-DE + MALDI sequencing : The differences in secretory phenotype * The development of novel therapeutic interventions in allergic disease{nameOfApplication=Show}
Proteomics which is clinical progression of Alzheimer's disease dementiaAlzheimer's disease dementia Clinic Problem Proteomics DiscussionAlzheimer's disease(AD)신경세포 사멸 신경원섬유 변화 노인반 시냅스 수의 감소 수상돌기 이상원인유전자 이상 면역학설 바이러스설유전자 이상신경원섬유 변화 노인반최근 연구프리세닐린 1과 2의 생물학적 기능 및 베타아밀로이드의 독성 기전 연구 AD에서 신경성장인자들의 역할및 상호관계 규명AD에 작용하는 신경교세포 마이크로리아의 연구 AD의 유전적 위험인자와 환경적 위험인자간의 상호 작용 연구AD의 진단CT(Computed Tomography) ScanAD의 치료아세틸콜린 전구체 아세틸콜린 분해 효소 억제제 칼슘 채널 브로커 호르몬 요법 신경펩타이드 모노아미 옥시다제-B 억제제 항염증 치료제문제점치료보다는 예방 Proteomics적인 방법이 필요MethodsSample 채취 Proteinchip arrays SELDI-TOF-MS Human genome-database cDNA 이식, 조직배양예상 결과신경 세포의 사멸 새로운 모델 필요DiscussionOn-line Two-dimensional Chromatography-Mass Spectrometry SEC + HPLC-MS{nameOfApplication=Show}
* Peptides play a central role in many physiological processes.* The use of technologies other than 2D gel in order to analyse all peptides and small proteins.* Using liquid chromatography or mass spectrometry have been developed and applied to the analysis of human body fluids.The establishment of human genome as the blueprint for human life - Necessary : the human proteome project → Understanding of the molecules that make up our complex regulatory systems Search for the biochemical messenger molecules - Regulatory processes : peptide hormones, neuropeptides, cytokines and enzyme inhibitors - Communication between all cell types through body fluids Significant progression in the field of proteomics - The sequences of secretin and insulin (the first examples) - Several important classes of peptide hormones : gut hormones, neuropeptides, leuteinizing hormone releasing hormone, growth hormonePeptides are of paramount importance ( Table 1 )The lack of technologies that enable a comprehensive analysis of peptides - Purifications from tissue extracts guided by function - Many steps are needed to generate a new sequence