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Hippocampal cell membrane proteomics and genomics dimensional chromatography on toxin research applications

Author: ZhouTieYang
Tutor: WangXianChun；LiangSongPing
School: Hunan Normal University
Course: Biochemistry and Molecular Biology
Keywords: Plasma membrane Proteomics Hippocampus Mass Electrophoresis Two-dimensional liquid Protein Spider toxin
CLC: Q51
Type: Master's thesis
Year: 2006
Downloads: 13
Quote: 0
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Abstract


Hippocampus (hippocampus) is the brain of a temporal lobe brain structure that is important for learning and memory mammalian structure area . Using differential centrifugation and discontinuous sucrose density gradient centrifugation purified adult rat hippocampus cell membrane and the plasma membrane by Western blotting on samples for testing. The results show that the sucrose density gradient centrifugation medium, the diaphragm focused on the sugar concentration of 32% to 42 % of the section ( density of about 1.13 to 1.18 ) , followed by 20% to 25% ( density of about 1.08 to 1.10 ) in section. 39% (density about 1.17 ) the level of the highest concentration and purity of the plasma membrane . One -dimensional SDS-PAGE separation and CapLC-MS/MS analysis, database searching through the cell membrane from rat hippocampal tissue samples were identified 135 kinds of proteins, membrane proteins and membrane-associated proteins with a total of 70 kinds ( 51.9% ) , including Na ~ / K ~-ATPase, Glutamate / aspartate transporter, Lipophilin, GLAST 1a and so on. To study the function of hippocampal tissue cells and rat hippocampus cell membrane protein database established and accumulated valuable information . China tiger tarantula is one of the most poisonous spiders , existing research shows that its venom is rich in low molecular weight (<1 kD) protein and peptide active ingredient. Isolation and identification of these components in order to establish a new two-dimensional liquid chromatography separation method . The method comprises on the one-dimensional isoelectric point (pI) and two-dimensional separation of the non-porous silica gel RP-HPLC separation . By ProteoVue and DeltaVue software will convert inverted spectrum with a similar two-dimensional gel electrophoresis images mimic diagram . Eluted component use Gilson 's collector to collect and use matrix-assisted laser desorption ionization - time of flight mass analysis. In the pI 3.5到9.0 1 ~ 10 kD molecular weight range of about 600 were detected bands of proteins and peptides , as measured by mass spectrometry analysis the molecular weight of component 130 , and with 26 the sequence information of the toxin components , including 12 kind of known toxins ingredients.

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CLC: > Biological Sciences > Biochemistry > Protein
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