Crystal structure and dimerization equilibria of PcoC, a methionine-rich copper resistance protein from Escherichia coli

PcoC is a soluble periplasmic protein encoded by the plasmid-born pco copper resistance operon of Escherichia coli. Like PcoA, a multicopper oxidase encoded in the same locus and its chromosomal homolog CueO, PcoC contains unusual methionine rich sequences. Although essential for copper resistance, the functions of PcoC, PcoA, and their conserved methionine-rich sequences are not known. Similar methionine motifs observed in eukaryotic copper transporters have been proposed to bind copper, but there are no precedents for such metal binding sites in structurally characterized proteins. The high-resolution structures of apo PcoC, determined for both the native and selenomethionine-containing proteins, reveal a seven-stranded beta barrel with the methionines unexpectedly housed on a solvent-exposed loop. Several potential metal-binding sites can be discerned by comparing the structures to spectroscopic data reported for copper-loaded PcoC. In the native structure, the methionine loop interacts with the same loop on a second molecule in the asymmetric unit. In the selenomethionine structure, the methionine loops are more exposed, forming hydrophobic patches on the protein surface. These two arrangements suggest that the methionine motifs might function in protein-protein interactions between PcoC molecules or with other methionine-rich proteins such as PcoA. Analytical ultracentrifugation data indicate that a weak monomer-dimer equilibrium exists in solution for the apo protein. Dimerization is significantly enhanced upon binding Cu(I) with a measured delta(deltaG degrees )相似文献   

Chemical and sensory characteristics of the fruits of eight cultivars of field grown tomato

The physical and sensory characteristics of tomato fruits (cf. Gajc-Wolska et al.: Physical and sensory characteristics of the fruits …, this issue) were supplemented by total sugars and titratable acid analysis and flavor. The instrumental and sensory methods were highly statistically correlated. Comparison of cultivars always requires that physical and chemical analysis of tomato fruit quality should always include results of sensory evaluation.     

Platinum(II) complexes with 2,4-diaminobutyric acid,ornithine, lysine and 4,5-diaminovaleric acid

L-2,4-Diaminobutyric acid (Dab) reacts with K₂PtCl₄ yielding PtCl₂(N,O-Dab), which rearranges to PtCl₂(N,N-Dab). Reaction with L-ornithine and L-lysine yields the corresponding PtCl₂(N,O-Orn) and PtCl₂(N,O-Lys), respectively, whereas reaction with 4,5-diaminovaleric acid (Dav) yields PtCl₂(N,N- Dav).     

Macronuclear differentiation in conjugating pairs of Tetrahymena treated with the antitubulin drug nocodazole

During Tetrahymena conjugation gamic nuclei (pronuclei) are produced, reciprocally exchanged, and fused in each mate. The synkaryon divides twice; the two anterior nuclei develop into new macronuclei while the two posterior nuclei become micronuclei. The postzygotic divisions were blocked with the antitubulin drug nocodazole (ND). Then pronuclei (gamic nuclei) developed directly into macronuclear anlagen (primordial macronuclei), inducing amicronucleate cells with two anlagen, or, rarely, cells with one anlagen and one micronucleus. ND had a similar effect on cells that passed the first postzygotic division inducing amicronucleate cells with two anlagen, while cells treated with ND at the synkarya stage produced only one large anlage. Different intracytoplasmic positioning of the nuclei treated with ND (pronuclei, synkarya and two products of the first division) shows that most of cell cytoplasm is competent for inducing macronuclear development. Only posteriorly positioned nuclei--products of the second postzygotic division--remain micronuclei. The total cell DNA content, measured cytophotometrically in control and in ND-induced amicronucleate conjugant cells with one and two anlagen, was similar in all three samples at 12 h of conjugation. Eventually, at 24 h this content was about 2 pg (8 C) per anlagen both in nonrefed control and in amicronucleate exconjugants. Therefore "large" nuclei developing in the presence of ND were true macronuclear anlagen.     

The effects of amphotericin B on lipid metabolism in cultured human skin fibroblasts

Summary Amphotericin B inhibits hydroxymethylglutarylcoenzyme A (HMGCoA) reductase activity and incorporation of [³H]acetate into sterols and fatty acids of human skin fibroblasts. Delivery of cholesterol to cells via the low density lipoprotein receptor pathway is not altered. The effects of amphotericin B on cell lipid metabolism are partially reversed by both potassium and insulin. Portions of this work were presented at the Southern Regional Meeting of the American Federation for Clinical Research, January 1983.     

Decline of Potentillo albae-Quercetum Libb. 1933 phytocoenoses in Poland

The historical process of the decline of the thermophilous oak forest was studied at 11 selected localities within the whole area of the association in Poland. The comparative analysis of phytosociological data from the periods of 1929–1981 and 1987–1991 has shown the total decline of Potentillo albae-Quercetum at nine localities as well as symptoms of regression at two localities. It has been stated that in Eastern Pomerania and in Greater Poland in the places of Potentillo albae-Quercetum phytocoenoses the communities of Galio-Carpinetum and Calamagrostio-Quercetum have appeared. In the central, north-east, south-east and southern part of the country, the thermophilous oak forest has changed into Tilio-Carpinetum phytocoenoses. The results of the study indicate that the recession of Potentillo albae-Quercetum is taking place in the whole area of the association in Poland. The size of the area is changing, the number of localities is decreasing. The consequence of the thermophilous oak forest decline is the unification and impoverishment of woodland vegetation and flora. These localities of the association which exist up to date are refuges for many interesting helio- and thermophilous species. Their presence to a great extent determines the genetic pool and diversity of Polish flora.Abbreviations Acc accompanying - FB Festuco-Brometea - Fg Fagetalia sylvaticae - MA Molinio-Arrhenatheretea - QF Querco-Fagetea - Qp Quercetalia pubescentis - SN Sedo-Scleranthetea and Nardo-Callunetea - TG Trifolio-Geranietea sanguinei - VP Vaccinio-Piceetea     

Comparison of Predictive Immunohistochemical Marker Expression of Primary Breast Cancer and Paired Distant Metastasis using Surgical Material: A Practice-Based Study

Parallel studies of primary breast carcinomas and corresponding distant metastases samples reveal considerable differences. Our aim was to highlight this issue from another perspective and provide further data based on 98 patient samples: 69 primary breast carcinoma and 85 distant metastases from bone, central nervous system (CNS) and lung (56 paired). Two independent series of immunohistochemical reactions with different antibodies for estrogen receptor (ER), progesterone receptor (PgR) and human epidermal growth factor receptor 2 (Her2), along with HER2 fluroscence in situ hybridization (FISH) were performed on tissue microarrays to classify breast carcinoma and distant metastases samples into Luminal A, Luminal B-proliferating, Luminal B-HER2+, HER2+ and triple negative (TNBC) surrogate breast cancer groups. Correlation and agreement between the two assessments of ER and PgR were fair-to-moderate, and almost perfect for HER2 and Ki67. There was 40% discordance concerning immunophenotype between breast carcinomas and distant metastases. Most common metastatic site of ER+ breast carcinoma was the skeletal system (59.2%), whereas that of TNBCs was the CNS (58.8%) and lungs (23.5%). Distant metastases in bones were mostly luminal (54.3%), in the CNS, Luminal B (53.2%), and in the lung, TNBC (37.5%). The change of drugable properties of primary breast cancers in the respective bone and CNS metastases suggests that characterization of the metastasis is necessary for appropriate treatment planning.