Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society最新文献_第5页

Characterization of gene expression in the cerebral cortices of rat brains containing subcortical lesions. 含有皮层下病变的大鼠脑皮层基因表达的表征。

Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society Pub Date : 1994-01-01

W Wallace, D Brane, N Hsu, N Khowong, C R Merril, V Haroutunian

{"title":"Characterization of gene expression in the cerebral cortices of rat brains containing subcortical lesions.","authors":"W Wallace, D Brane, N Hsu, N Khowong, C R Merril, V Haroutunian","doi":"","DOIUrl":"","url":null,"abstract":"Neurotoxic lesion of the nucleus basalis of Meynert in the rat brain, which results in the loss of subcortical cholinergic innervation to the cerebral cortex, is an animal model for the cortical cholinergic deficits that are characteristic of Alzheimer's disease. Previously, we have shown that amyloid precursor protein is induced in the cortex in response to this disrupted innervation. We have investigated the synthesis and accumulation of proteins in lesioned versus control cortices. Total proteins from cortices were separated by high resolution two-dimensional gel electrophoresis and visualized by silver stain. Of the greater than 1,000 polypeptides examined, only one exhibited a consistent alteration in the lesioned sample. This unidentified protein (Mr 34 kD, pI 5.5) was normally present in scant amounts but was virtually absent in the lesioned cortex (0.056% total integrated density (TID) and 0.008% TID, respectively; p < 0.04). To investigate gene expression more directly, polysomes purified from lesioned and control cortices were assayed in vitro. Examination of [35S] incorporation into translation products by two-dimensional gels and autoradiography revealed three newly synthesized polypeptide differences in the lesioned samples. One protein (M(r) 47 kD, pI 6.1) exhibited elevated levels with the lesion (0.05% to 0.16%; p = 0.02) while two other proteins (M(r) 34 kD, pI 5.5, and M(r) 33 kD, pI 5.7) exhibited reduced levels (0.20% to 0.04%, p < 0.02, and 0.34% to 0.12%, p = 0.04, respectively).","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"4 1","pages":"33-8"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18810973","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

On the "door-corridor" model of gel electrophoresis. III. The gel constant and resistance, and the net charge, friction, diffusion and electrokinetic force of the migrating molecules. 关于凝胶电泳的“门-走廊”模型。3凝胶常数和阻力，以及迁移分子的净电荷、摩擦、扩散和电动势。

Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society Pub Date : 1994-01-01

B Kozulić

{"title":"On the \"door-corridor\" model of gel electrophoresis. III. The gel constant and resistance, and the net charge, friction, diffusion and electrokinetic force of the migrating molecules.","authors":"B Kozulić","doi":"","DOIUrl":"","url":null,"abstract":"The door-corridor model of gel electrophoresis enabled an estimation of the net charge of DNA molecules run in various gels. When the runs were carried out in Trisacetate-EDTA buffer having a concentration from 10 to 120 mM, the net charge in 1% agarose gel varied from 1.1 to 0.58 e per base pair. The friction between migrating molecules and gel fibers was dependent on the gel type and concentration, as well as the electric field strength and temperature during electrophoresis. In the 123 to 1,474 bp size range, the friction in 1% agarose changed from 1.91 to 378.03 x 10(-10) N.m-1.s. It was found that the friction per 123 bp DNA segment is not constant, but raises with size. The gel resistance force increases at higher electric field strengths, indicating that elastic forces govern the migration of macromolecules through gels. In the gels studied, the friction, and therefore thermal diffusion, of DNA and protein-SDS complexes scale with from 2.20 to 2.32 power of size. The ratio of thermally induced diffusion and velocity in various gels shows that there is a profound reduction of diffusion compared to velocity with increasing DNA size. This is directly linked to the high exponent relating friction and size. The high resolving power of gel electrophoresis can be correlated to the difference between the frictional coefficients of a diffusing and migrating macromolecule.","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"4 3","pages":"149-59"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18616766","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Mouse cells with null p53 mutation have all p53 isoforms deleted and lose negative growth control. 缺失p53突变的小鼠细胞缺失了所有p53同工型，失去了负生长控制。

Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society Pub Date : 1994-01-01

J K Selkirk, C He, B A Merrick

引用次数: 0

On the "door-corridor" model of gel electrophoresis. I. Equations describing the relationship between mobility and size of DNA fragments and protein-SDS complexes. 关于凝胶电泳的“门-走廊”模型。描述DNA片段和蛋白质- sds复合物的迁移率和大小之间关系的方程。

Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society Pub Date : 1994-01-01

B Kozulić

{"title":"On the \"door-corridor\" model of gel electrophoresis. I. Equations describing the relationship between mobility and size of DNA fragments and protein-SDS complexes.","authors":"B Kozulić","doi":"","DOIUrl":"","url":null,"abstract":"In a gel, electrophoretic mobility of a DNA fragment and a protein-SDS complex is an exponential function of the ratio between two forces, the resisting force of the gel polymers and the electrokinetic force of the migrating macromolecule. It is also proportional to the mobility of unit size of the migrating molecule, mu 1. Each gel gives an optimal resolution of macromolecules of a defined size. That size must be such that the electrokinetic force of the migrating molecule equals the gel resistance force, so that the ratio of mobility and mu 1 becomes e-1. The optima for several poly[N-acryloyl-tris(hydroxymethyl)aminomethane], polyacrylamide and agarose gels are given. Only two constants are sufficient for description of the relationship between mobility and size of DNA fragments and protein-SDS complexes of the sizes close to that optimally resolved. One is mu 1 and the other, ks, is the slope of the straight line which can be obtained from the size versus reciprocal of the mobility plot. The experimentally observed sigmoidal curves in the plots including the logarithm of size follow directly from the developed equations. The use of mu 1 instead of mu 0 may provide a better interpretation of the Ferguson plots.","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"4 3","pages":"125-36"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18619479","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Multiple p53 protein isoforms and formation of oligomeric complexes with heat shock proteins Hsp70 and Hsp90 in the human mammary tumor, T47D, cell line. 人乳腺肿瘤T47D细胞系中多种p53蛋白同工型及与热休克蛋白Hsp70和Hsp90寡聚复合物的形成

Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society Pub Date : 1994-01-01

J K Selkirk, B A Merrick, B L Stackhouse, C He

{"title":"Multiple p53 protein isoforms and formation of oligomeric complexes with heat shock proteins Hsp70 and Hsp90 in the human mammary tumor, T47D, cell line.","authors":"J K Selkirk, B A Merrick, B L Stackhouse, C He","doi":"","DOIUrl":"","url":null,"abstract":"At least eleven isoforms of p53 protein were observed in a human mammary tumor cell line. T47D. Comparative 33P and 35S incorporation analysis showed an equal distribution of P53 isoforms within cytoplasmic and nuclear compartments, although phosphorylation was unequal among isoforms and the most basic p53 species was unphosphorylated. Using a combination of immunoprecipitation with monoclonal antibodies for p53 and heat shock proteins Hsp70 & Hsp90, and two-dimensional gel electrophoretic analysis, T47D p53 protein oligomers were observed with several species of Hsp70 and Hsp90. The p53/Hsp70/Hsp90 aggregate dissociates after nuclear translocation. Immunoprecipitation of Hsp70 and Hsp90 using monoclonal antibodies showed formation of a heteroligomer between Hsp70 and Hsp90 in cytoplasm but not nucleus. This suggests these Hsp proteins can form a complex in the cytoplasm but undergo a conformational change after nuclear translocation such that Hsp/Hsp binding sites are no longer recognized. These data indicate T47D cells have multiple p53 precursor molecules probably at different stages of phosphorylation, and which may be sequestered from proteases by binding to Hsp proteins. Hsp proteins also can heterocomplex in the cytoplasm, also possibly as protection against protease degradation until bound to p53. After translocation, p53 is freed from Hsp proteins for binding to DNA where Hsp70 and Hsp90 are no longer able to form a nuclear complex probably rendering Hsp's labile to proteolysis.","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"4 1","pages":"11-8"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18810969","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A classification of one-dimensional electrophoresis gels using wave packet decomposition. 用波包分解对一维电泳凝胶进行分类。

Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society Pub Date : 1994-01-01

A Zahnd, M Funk, P Vaudaux, D Lew, J R Scherrer, D F Hochstrasser

引用次数: 0

Brief report: electrophoretic separation of oligonucleotides in replenishable polyacrylamide-filled capillaries. 简要报告:可补充聚丙烯酰胺填充的毛细血管中寡核苷酸的电泳分离。

Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society Pub Date : 1994-01-01

C Heller, J L Viovy

引用次数: 0

Electrophoretic free mobility and viability of microbial cells: a preliminary study in preparation for space experiments. 微生物细胞的电泳自由迁移率和活力:为空间实验做准备的初步研究。

Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society Pub Date : 1994-01-01

T Akiba, A Nishi, M Takaoki, S Nagaoka, F Tomita

引用次数: 0

A model of gel electrophoresis. 凝胶电泳模型。

Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society Pub Date : 1994-01-01

B Kozulić

引用次数: 0

Two-dimensional gel electrophoresis of major cytosolic proteins derived from spleen mononuclear cells of normal and leukemic rats. 正常和白血病大鼠脾单核细胞主要胞质蛋白的二维凝胶电泳。

Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society Pub Date : 1993-01-01

B A Merrick, C He, M P Dieter, J K Selkirk

{"title":"Two-dimensional gel electrophoresis of major cytosolic proteins derived from spleen mononuclear cells of normal and leukemic rats.","authors":"B A Merrick, C He, M P Dieter, J K Selkirk","doi":"","DOIUrl":"","url":null,"abstract":"Proteins from leukemic spleen cells derived from a F344 rat transplant model were investigated by 2D gel electrophoresis. The cell line was maintained in vivo by serial transplant of mononuclear spleen cells from leukemic donors into syngeneic recipients. Cytosolic proteins from mononuclear cells (MNC) were isolated from spleens of normal and leukemic rats and separated by electrophoresis. Replicate data from silver-stained gels for each preparation were compiled into Master Images using image analysis algorithms in order to characterize the normal and leukemic protein profiles. Comparative analysis showed a total of 458 proteins that were reproducibly detected in normal MNC, while almost twice the number of proteins (828) were found in leukemic MNC, suggestive of the more mitotically active tumor cells. Profile analysis showed that normal and leukemic preparations shared 228 common proteins, with 600 proteins observed only in leukemic MNC, and 230 proteins found only in normal MNC. Differences in protein patterns between normal and leukemic MNC in rats probably reflects a shift in spleen leukocyte populations and a relative induction of gene expression in leukemic MNC as compared to the normal MNC.","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"3 5","pages":"203-11"},"PeriodicalIF":0.0,"publicationDate":"1993-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19207938","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0