摘要:

AbstractA total of 3430 polypeptides (2592 cellular; 838 secreted) from transformed human amnion cells (AMA) labeled with [35S]methionine were separated and recorded using computer‐aided two‐dimensional (2‐D) gel electrophoresis. A master 2‐D gel database of cellular protein information that includes both qualitative and quantitative annotations has been established. The protein numbers in this database differ from those reported in an earlier version (Celiset al. Leukemia1988,2, 561–602) as a result of changes in the scanning hardware. The reported information includes: percentage of total radioactivity recovered from the gels (based on quantitations of polypeptides labeled with a mixture of 1614C‐amino acids), protein name (including credit to investigators that aided identification), antibody against protein, cellular localization, (nuclear, 40S hnRNP, 20S snRNP U5, proteasomes, endoplasmic reticulum, mitochondria, Golgi, ribosomes, intermediate filaments, microfilaments and microtubules), levels in fetal human tissues, partial protein sequences (containing information on 48 human proteins microsequenced so far), cell cycle‐regulated proteins, proteins sensitive to interferons α, β, and γ, heat shock proteins, annexins and phosphorylated proteins. The results presented should be considered as the initial phase of a joint effort between our laboratories to undertake a general and systematic analysis of human proteins. Using this integrated approach it will be possible to identify phenotype‐specific proteins, to microsequence them and store the information in the database, to identify the corresponding genes, to search for homology with previously characterized proteins and to study the function of groups of proteins (pathways, organelles,etc.) that exhibit interesting regulatory properties. In particular, the 2‐D gel protein database may become increasingly important in view of the concerted effort to map and sequence the

ISSN:0173-0835    

DOI:10.1002/elps.1150111202

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

年代:1990

数据来源: WILEY

摘要:

AbstractA new version of the MRC‐5 two‐dimensional gel cellular protein database (Celiset al., Electrophoresis1989,10, 76–115) is presented. Gels were scanned with a Molecular Dynamics laser scanner and processed by the PDQUEST IITMsoftware. A total of 1895 [35S]methionine‐labeled cellular polypeptides (1323 with isoelectric focusing and 572 with nonequilibrium pH gradient electrophoresis) are recorded in this database, containing quantitative and qualitative data on the relative abundance of cellular proteins synthesized by quiescent, proliferating and SV40 transformed MRC‐5 fibroblasts. Of the 592 proteins quantitated so far, the levels of 138 were up‐or down‐ regulated (51 and 87, respectively) by two times or more in the transformed cells as compared to their normal proliferating counterparts, while only 14 behaved similarly in quiescent cells. Seven MRC‐5 SV40 proteins, including plastin and two interferon‐induced proteins, were not detected in the master MRC‐5 images. The identity of 36 of the transformation‐sensitive proteins whose levels are up or down regulated by two times or more was determined and additional information can be transferred from the master transformed human epithelial amnion cells (AMA) database (Celiset al., Electrophoresis1990,11, 989–1071) for those polypeptides of known and unknown identity that have been matched to AMA polypeptides. As more information is gathered in this and other laboratories, including data on oncogene proteins and transcription factors, this comprehensive database will outline an integrated picture of the expression levels and properties of the thousands of protein components of organelles, pathways and cytoskeletal systems that may be directly or indirectly involved in properties associated wit

ISSN:0173-0835    

DOI:10.1002/elps.1150111203

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

年代:1990

数据来源: WILEY

摘要:

AbstractQuantitative protein databases reveal the response of cells to experimental variables, such as exposure to growth factors or transfection with a transforming gene. The nature of the response depends on the type of cell and its internal state at the time of the stimulus. By constructing a protein database to study a given cell line, we can better understand the differentiated state of the cell, the growth regulatory mechanisms it employs, the particular mechanisms it uses to cope with its environment, and the ways these mechanisms may have been compromised through mutation or transformation. The REF52 database is a quantitative database designed to study growth control and transformation in a well‐defined family of normal and transformed rat cell lines. The database, which has been described and analyzed elsewhere (J. I. Garrels and B. R. Franza,J. Biol. Chem.1989,264, 5283–5298 and J. I. Garrels and B. R. Franza,J. Biol. Chem.1989,264, 5299–5312) is further explored here using cluster analysis. This method reveals the most common protein expression profiles for each series of two‐dimensional gels without requiring any prior hypothesis or queries on the part of the investigator. This study reveals, for each experiment, large and small clusters of protein expression profiles, most of which have readily apparent biological meaning. For example, large clusters of proteins induced or repressed during growth to confluence have been revealed, and several clusters of transformation‐sensitive proteins reveal differential effects of transformation by DNA‐ and RNA‐tumor viruses. This analysis extends our earlier quantitative explorations of the REF52 protein database and helps to show how such a database can be used to provide context and guidance for molecular studies of regulation in a give

ISSN:0173-0835    

DOI:10.1002/elps.1150111204

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

年代:1990

数据来源: WILEY

摘要:

AbstractThe first two editions of theE. coliGene‐Protein Index were published to provide identifications of protein spots resolved by two‐dimensional gel electrophoresis as the products of known genes. This third edition has been expanded to include information about genes and proteins gained directly from two‐dimensional gel analysis – including information about protein spots not yet characterized genetically or biochemically – and is therefore more properly called a cellular proteins database. An alpha‐numeric designation has been uniquely assigned to each of the 616 polypeptide spots in the current database. To this, information is linked about the polypeptide′s identification (protein name, gene name, Enzyme Commission – EC number), location on reference gels (x‐ycoordinates), genetics (Genbank code, DNA sequence reference), biochemistry (molecular weight, isoelectric point), and physiology (steady state level of the protein as a function of media and temperature, membership in various regulo

ISSN:0173-0835    

DOI:10.1002/elps.1150111205

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

年代:1990

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.1150111206

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

年代:1990

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.1150111208

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

年代:1990

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.1150111201

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

年代:1990

数据来源: WILEY