ISSN:0173-0835    

DOI:10.1002/elps.1150100202

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1989

数据来源: WILEY

摘要:

AbstractA workshop entitled Cellular Protein Databases from Two‐Dimensional Gel Electrophoresis was held in Atlanta, Georgia, 28 February–1 March 1987. Its purpose was to assess the status of two‐dimensional gel electrophoresis of proteins as a research methodology in biological systems, particularly in the generation of cellular protein databases. The workshop participants summarized current studies on a variety of biological systems, both prokaryotic and eukaryotic. Analysis of the progress being made led to the conclusion that electrophoretic techniques, supported by automatic scanning of gel images and computer‐assisted processing, analysis and matching of gel images, are now capable of generating databases of great potential value. Factors affecting the reproducibility of protein spot patterns on gels were identified, and the extent to which gel pattern variability causes difficulties in communicating results and in integrating information from different laboratories was assessed. Measures were suggested to help overcome obstacles to the generation of comprehensive cellular protein databases from the electrophoretic resolution of total cellular p

ISSN:0173-0835    

DOI:10.1002/elps.1150100203

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1989

数据来源: WILEY

摘要:

AbstractDatabases of protein information from human embryonal lung fibroblasts (MRC‐5) have been established using computer analyzed two‐dimensional gel electrophoresis. One thousand four hundred and eighty‐two cellular proteins (1060 with isoelectric focusing and 422 with nonequilibrium pH gradient electrophoresis, in the first dimension) ranging in molecular mass between 8 and 234 kDa were separated and numbered. Information entered in the database (in most cases for major proteins) includes: protein name, HeLa protein catalog number, mouse protein catalog number, proteins matched in transformed human epithelial amnion cells (AMA) and peripheral blood mononuclear cells (PBMC), transformation and/or proliferation sensitive proteins, synthesis in quiescent cells, cell cycle regulated proteins, mitochondrial and heat shock proteins, cytoskeletal proteins and proteins whose synthesis is affected by interferons. Additional information entered for a few transformation‐sensitive proteins that have been selected for future studies includes levels of synthesis and amounts in fetal human tissues. A total of four hundred and seventy‐six [35S] methionine labeled polypeptides (258 isoelectric focusing; 218, nonequilibrium pH gradient electrophoresis) secreted by MRC‐5 fibroblasts were separated and recorded (J. E. Celiset al., Leukemia1987,1, 707–717). Information entered in this database includes molecular weight and transformation sensitive proteins. These databases, as well as those of epithelial and lymphoid cell proteins (J. E. Celiset al., Leukemia1988,9, 561–601), represent the initial stages of a systematic effort to establish comprehensive databases of human protein information. In the long run, these databases are expected to offer a useful framework in which to focus the human genome se

ISSN:0173-0835    

DOI:10.1002/elps.1150100204

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1989

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.1150100205

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1989

数据来源: WILEY

摘要:

AbstractIn its most useful form a cellular protein database should be genomically based, because it is the genome which determines both the total number of proteins a cell can make and the particular ones that will be made under any given condition. Such a database should trace each protein back to its structural gene, and should account for every structural gene of a cell. Recent advances in molecular biology greatly facilitate the construction of such gene‐protein databases. The mapping of genes of unidentified proteins resolved from total cell extracts on two‐dimensional gels can now be accomplished by largely biochemical methods, without the necessity of isolating mutants or performing genetic crosses. Other techniques permit one to search gels for the product of any newly discovered gene (or open reading frame) suspected of encoding a protein. Consequently, gene‐protein indices can be built independently and simultaneously from either direction – deducing the genetic map from the protein pattern, or finding the protein pattern from information encoded in the genome. A database of this sort is being constructed for the bacterium,Escherichia coli. Given the current pace of DNA nucleotide sequencing, the development of total gene‐protein indices for a variety of cells can be anticipated in the ne

ISSN:0173-0835    

DOI:10.1002/elps.1150100206

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1989

数据来源: WILEY

摘要:

AbstractTwo‐dimensional (2‐D) polyacrylamide gel electrophoresis can detect thousands of polypeptides, separating them by apparent molecular weight (Mr) and isoelectric point (pI). Thus it provides a more realistic and global view of cellular genetic expression than any other technique. This technique has been useful for finding sets of key proteins of biological significance. However, a typical experiment with more than a few gels often results in an unwieldy data management problem. In this paper, the GELLAB‐II system is discussed with respect to how data reduction and exploratory data analysis can be aided by computer data management and statistical search techniques. By encoding the gel patterns in a “three‐dimensional” (3‐D) database, an exploratory data analysis can be carried out in an environment that might be called a “spread sheet for 2‐D gel protein data”. From such databases, complex parametric network models of protein expression during events such as differentiation might be constructed. For this, 2‐D gel databases must be able to include data from other domains external to the gel itself. Because of the increasing complexity of such databases, new tools are required to help manage this complexity. Two such tools, object‐oriented databases and expert‐system rule‐based analysis, are discussed in this context. Comparisons are made between GELLAB and other 2‐D gel databases analysis systems to illustrate some of the analysis paradigms common to these systems and where t

ISSN:0173-0835    

DOI:10.1002/elps.1150100207

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1989

数据来源: WILEY

摘要:

AbstractHigh‐resolution two‐dimensional electrophoresis (2‐DE) has reached a technological level that allows us to resolve most of the numerous unknown protein species of a mammalian organism if appropriate strategies are used. We will discuss the problems of classification and characterization of proteins and propose a systematic approach to the analysis of the total protein complex. Both a comprehensive as well as a pragmatic approach towards systematic analysis have been considered. A “complex protein database” is suggested and considered with regard to various uses. A systematic analysis of the mouse proteins has been started and some of the preliminary results are summarized here. In particular, genetic properties of the proteins were investigated and are presented in order to demonstrate the significance of a systematic analysis of proteins for research and practical application (e. g.mutagenicity testing). A concept is presented for sequencing the coding DNA of mouse and man, starting with a systematic analysis of mouse proteins and then using two recently developed methods ‐ microsequencing of proteins from spots of 2‐DE protein patterns, and utilization of the relatively shortN‐terminal sequences obtained – to produce the corresponding cDNA's

ISSN:0173-0835    

DOI:10.1002/elps.1150100208

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1989

数据来源: WILEY

摘要:

AbstractAs part of our continuing studies into the biochemical basis of long‐term changes in neuronal function inAplysia, we have developed a simple method for obtaining amino acid sequence information from proteins isolated on two‐dimensional gels. Proteins isolated on preparative two‐dimensional gels are digestedin situwithStaphylococcus, aureusV8 protease, and the resulting peptides electrophoresed, transferred to a polyvinylidene difluoride membrane, and sequenced in a gas‐phase sequencer. The method is simple and should be applicable to a variety of other systems where the development of a two‐dimensional gel database is

ISSN:0173-0835    

DOI:10.1002/elps.1150100209

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1989

数据来源: WILEY

摘要:

AbstractIn this paper, techniques are demonstrated for unambiguously mapping phosphoproteins and glycoproteins into a protein database of unfractionated fibroblast proteins. These methods allow for precise registration of subsets of proteins with the image of total proteins. The methods are also applicable to database mapping of antigens identified by indirect immunodetection methods and receptor molecules identified with photoaffinity ligands.

ISSN:0173-0835    

DOI:10.1002/elps.1150100210

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1989

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.1150100211

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1989

数据来源: WILEY