A standard two-dimensional (2-D) protein map of Fischer 344 rat liver (F344MST3) is presented, with a tabular listing of more than 1200 protein species. Sodium dodecyl sulfate (SDS) molecular mass and isoelectric point have been established, based on positions of numerous internal standards. This map has been used to connect and compare hundreds of 2-D gels of rat liver samples from a variety of studies, and forms the nucleus of an expanding database describing rat liver proteins and their regulation by various drugs and toxic agents. An example of such a study, involving regulation of cholesterol synthesis by cholesterol-lowering drugs and a high-cholesterol diet, is presented. Since the map has been obtained with a widely used and highly reproducible 2-D gel system (the Iso-Dalt system), it can be directly related to an expanding body of work in other laboratories.

We have improved upon the reference two-dimensional (2-D) electrophoretic map of rat liver proteins originally published in 1991 (N. L. Anderson et al., Electrophoresis 1991, 12, 907-930). A total of 53 proteins (102 spots) are now identified, many by microsequencing. In most cases, spots cut from wet, Coomassie Blue stained 2-D gels were submitted to internal tryptic digestion [2], and individual peptides, separated by high-performance liquid chromatography (HPLC), were sequenced using a Perkin-Elmer 477A sequenator. Additional spots were identified using specific antibodies.

Two-dimensional gel electrophoresis of denaturated proteins were performed at five developmental stages or organs (hereafter referred to as stage-organs) on two wheat lines with four different cytoplasms. Five hundred and fifty to 712 reproducible spots were scored depending on the stage-organ. Each stage-organ is unambiguously characterized and several types of control of protein quantity are recorded. Post-translational modifications are hypothetized and may sometimes be stagespecific. Two cytoplasmic patterns are found: one for the euplasmic lines with Triticum aestivum cytoplasm and one for the alloplasmic lines with Aegilops juvenalis, Ae. ventricosa and Ae. kotschyi cytoplasms. Cytoplasmic variation is observed for 28 spots showing position difference, all of which are probably products of the LS gene, and for four spots showing differences for regulation of protein quantity. Nuclear variation between 'Chinese Spring' and 'Selkirk' is found for 20 allelic differences and for 20 regulatory systems, the latter number being probably underestimated.

In this first analysis the protein patterns obtained by two-dimensional gel electrophoresis of 8 day-old leaves from 18 alloplasmic wheat lines are compared. From 440 spots retained on the basis of their reproducibility, 36 proteins were observed to vary in different cytoplasms, allowing us to distinguish the T. aestivum cytoplasm from 5 Aegilops cytoplasms. Twenty-four of the 36 variable proteins could be structurally related to the large subunit of RuBPCase. Nuclear variation between 3 wheat varieties was observed for 14 proteins.

The changes in thrombocyte proteins during 22 degrees C storage of platelet concentrates (PC) were studied. To prepare a reference protein "map" of stored PC, platelet samples were taken on days 1, 7, and 21. The platelet proteins were separated by isoelectric focusing (first-dimension) followed by second-dimension polyacrylamide gradient gel electrophoresis with sodium dodecylsulfate (2D). The silver-stained gels were analyzed by computer to obtain a composite map of stored PC proteins. The pattern seen on day 1 changed during 7 days of storage, with 30 proteins increasing or decreasing in spot density. In general, the spot density for lower-molecular-weight proteins increased, whereas that for higher-molecular-weight proteins decreased. Membrane proteins of intact fresh and stored platelets were labeled with 3H using sodium metaperiodate-[3H]NaBH4 and with 125I using lactoperoxidase-H2O2. A comparison on the fluorographs of 2D gels of [3H]NaBH4-labeled platelet proteins showed several protein spots in the stored Day 7 sample that had not been seen in the Day 1 sample. Similarly, for the autoradiographs, several 125I-labeled proteins detected in Day 7 PC were not seen in the Day 1 samples. The results provide evidence that platelet proteins are altered during PC storage and that these changes involve platelet membrane proteins.