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Cytosolic oestrogen receptor content of breast cancer tissue in blacks and whites.: South African medical journal = Suid-Afrikaans e tydskrif vir geneeskunde, Vol. 60, No. 6. (8 August 1981)We have examined the oestradiol receptor (ERc) content of cytosols from 560 Black and White patients. The tumours from the White group contained a significantly lower proportion of ERc- negative tumours (35.4%) compared with those from the Black group (46.5%). The proportion of ERc-positive tumours is significantly greater in the post- than in the premenopausal White group. The frequencies of ERc-positive tumours in the post- and premenopausal Black groups are not significantly different, resembling the situation described by others in Japanese women.RJ Pegoraro, SM Joubert

Source: South African medical journal = Suid-Afrikaanse tydskrif vir geneeskunde, Vol. 60, No. 6. (8 August 1981)
Mass spectrometric analysis of rat ovary and testis cytosolic glutathione S-transferases (GSTs): identification of a novel class-alpha GST, rGSTA6*, in rat testis.: Biochem J, Vol. 323 ( Pt 2) (15 April 1997), pp. 503-510.Cytoso lic glutathione S-transferases (GSTs) from rat ovaries and testis were purified by a combination of GSH and S-hexylglutath ione affinity chromatography . The isolated GSTs were subjected to reverse-phase HPLC, electrospray MS and N-terminal peptide sequencing analysis. The major GST isoenzymes expressed in ovaries are subunits A3, A4, M1, M2 and P1. Other isoenzymes detected are subunits A1, M3 and M6*. In rat testis, the major GST isoenzymes expressed are subunits A3, M1, M2, M3, M5* and M6*. Subunits A1, A4 and P1 are expressed in lesser amounts. We could not detect post-translati onal modifications of any GSTs with known cDNA sequence. The molecular masses of subunits M5* and M6*, two class-Mu GSTs that have not been cloned, were determined to be 25495 and 26538 Da respectively. An N-terminally modified protein from rat testis with molecular mass 25737 Da was isolated from the S-hexylglutath ione column. Results from internal peptide sequencing analysis indicate that this is a novel class-Alpha GST that has not been previously reported. We designate this protein rGSTA6*.CH Hsieh, SP Tsai, HI Yeh, TC Sheu, MF Tam

Source: Biochem J, Vol. 323 ( Pt 2) (15 April 1997), pp. 503-510.
Effects of solution crowding on actin polymerization reveal the energetic basis for nucleotide-dep endent filament stability: Journal of Molecular Biology, Vol. In Press, Accepted ManuscriptKend ra Frederick, David Sept, De La

Source: Journal of Molecular Biology, Vol. In Press, Accepted Manuscript
The Redox Environment in the Mitochondrial Intermembrane Space Is Maintained Separately from the Cytosol and Matrix: J. Biol. Chem., Vol. 283, No. 43. (24 October 2008), pp. 29126-29134.Re dox control in the mitochondrion is essential for the proper functioning of this organelle. Disruption of mitochondrial redox processes contributes to a host of human disorders, including cancer, neurodegenerat ive diseases, and aging. To better characterize redox control pathways in this organelle, we have targeted a green fluorescent protein-based redox sensor to the intermembrane space (IMS) and matrix of yeast mitochondria. This approach allows us to separately monitor the redox state of the matrix and the IMS, providing a more detailed picture of redox processes in these two compartments. To verify that the sensors respond to localized glutathione (GSH) redox changes, we have genetically manipulated the subcellular redox state using oxidized GSH (GSSG) reductase localization mutants. These studies indicate that redox control in the cytosol and matrix are maintained separately by cytosolic and mitochondrial isoforms of GSSG reductase. Our studies also demonstrate that the mitochondrial IMS is considerably more oxidizing than the cytosol and mitochondrial matrix and is not directly influenced by endogenous GSSG reductase activity. These redox measurements are used to predict the oxidation state of thiol-containi ng proteins that are imported into the IMS. 10.1074/jbc.M8 03028200Jingji ng Hu, Lixue Dong, Caryn Outten

Source: J. Biol. Chem., Vol. 283, No. 43. (24 October 2008), pp. 29126-29134.
Insight into the proteome of the hyperthermophi lic Crenarchaeon Ignicoccus hospitalis: the major cytosolic and membrane proteins: Archives of Microbiology, Vol. 190, No. 3. (27 June 2008), pp. 379-394.Ignico ccus hospitalis, a hyperthermophi lic, chemolithoauto trophic Crenarchaeon, is the host of Nanoarchaeum equitans. Together, they form an intimate association, the first among Archaea. Membranes are of fundamental importance for the interaction of I. hospitalis and N. equitans, as they harbour the proteins necessary for the transport of macromolecules like lipids, amino acids, and cofactors between these organisms. Here, we investigated the protein inventory of I. hospitalis cells, and were able to identify 20 proteins in total. Experimental evidence and predictions let us conclude that 11 are soluble cytosolic proteins, eight membrane or membrane-assoc iated proteins, and a single one extracellular. The quantitatively dominating proteins in the cytoplasm (peroxiredoxin ; thermosome) antagonize oxidative and temperature stress which I. hospitalis cells are exposed to at optimal growth conditions. Three abundant membrane protein complexes are found: the major protein of the outer membrane, which might protect the cell against the hostile environment, forms oligomeric complexes with pores of unknown selectivity; two other complexes of the cytoplasmic membrane, the hydrogenase and the ATP synthase, play a key role in energy production and conversion.Til lmann Burghardt, Manfred Saller, Sonja Gurster, Daniel Muller, Carolin Meyer, Ulrike Jahn, Eduard Hochmuth, Rainer Deutzmann, Frank Siedler, Patrick Babinger, Reinhard Wirth, Harald Huber, Reinhard Rachel

Source: Archives of Microbiology, Vol. 190, No. 3. (27 June 2008), pp. 379-394.
Structural organization of interphase 3T3 fibroblasts studied by total internal reflection fluorescence microscopy.: J Cell Biol, Vol. 100, No. 4. (April 1985), pp. 1091-1102.We studied the laminar organization of 3T3 fibroblast cells growing on glass slides by use of total internal reflection illumination to excite fluorescence emission (TIRF) from labeled molecules and stained cellular compartments that are very close to the cell-substrate contact region. Mitochondria, distant from the contact regions and stained with the water-soluble cationic dye, dil-C3-(3), fluoresced only as the glass/cytoplas m critical angle was approached. A similar result was obtained when the nuclei were stained with Hoechst dye 33342. From this measured angle a cytoplasmic refractive index in the range 1.358-1.374 was computed. The plasma membrane of 3T3 cells was stained with dil-C18-(3), and the cytoplasmic compartment was stained with fluoresceinyl- dextran (FTC-dextran) or with carboxyfluores cein. We have demonstrated a high degree of correspondence between the low-reflectanc e zones in the reflection interference image of a live cell and the TIRF images of both the plasma membrane and cytoplasmic compartment. TIRF photometry of selected contact regions of cells provided data from which the absolute separation of cell and substrate was computed. From a population of 3T3 cells microinjected with fluorescein-la beled actin, motile and adherent interphase cells were selected for study. For adherent cells, which displayed fluorescent stress fibers, the TIRF image was composed of intense patches and less intense regions that corresponded, respectively, to the focal contact and close-contact zones of the reflection-int erference image. The intense patches corresponded to the endpoints of the stress fibers. Cells of motile morphology, which formed some focal contacts and extensive close-contact zones, gave AF-actin TIRF images of relatively even intensity. Thin lamellar regions of the cytoplasm were found to contain concentrations of actin not significantly different from other close-contact regions of the cell. The major analytical problem of TIRF microscopy is separation of the effects of proximity to substrate, refractive index, and fluorescent probe concentration on the local brightness of the TIRF image. From our results, it appears possible to use TIRF microscopy to measure the proximity of different components of substrate contact regions of cells.F Lanni, AS Waggoner, DL Taylor

Source: J Cell Biol, Vol. 100, No. 4. (April 1985), pp. 1091-1102.
Measurement of cytosolic Ca2+ concentration in Limulus ventral photoreceptors using fluorescent dyes: J. Gen. Physiol., Vol. 105, No. 1. (1 January 1995), pp. 95-116.10.1085 /jgp.105.1.95K Y Ukhanov, TM Flores, HS Hsiao, P Mohapatra, CH Pitts, R Payne

Source: J. Gen. Physiol., Vol. 105, No. 1. (1 January 1995), pp. 95-116.

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