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91.
Helen Pappa William R. Patterson T. L. Poulos 《Journal of biological inorganic chemistry》1996,1(1):61-66
The crystal structures of ascorbate peroxidase (APX) and cytochrome c peroxidase (CCP) show that the active site structures are nearly identical. Both enzymes contain a His-Asp-Trp catalytic triad in the proximal pocket. The proximal Asp residue hydrogen bonds with both the His proximal heme ligand and the indole ring nitrogen of the proximal Trp. The Trp is stacked parallel to and in contact with the proximal His ligand. This Trp is known to be the site of free radical formation in CCP compound I and also is essential for activity. However, APX forms a porphyrin radical and not a Trp-centered radical, even though the His-Asp-Trp triad structure is the same in both peroxidases. We found that conversion of the proximal Trp to Phe has no effect on APX enzyme activity and that the mutant crystal structure shows that changes in the structure are confined to the site of mutation. This indicates that the paths of electron transfer in CCP and APX are distinctly different. The Trp-to-Phe mutant does alter the stability of the APX compound I porphyrin radical, by a factor of two. Electrostatic calculations and modeling studies show that a potassium cation located about 8?Å from the proximal Trp in APX, but absent in CCP, makes a significant contribution to the stability of a cation Trp radical. This underscores the importance of long-range electrostatic effects in enzyme catalyzed reactions. 相似文献
92.
Further progress towards a catalogue of all Arabidopsis genes: analysis of a set of 5000 non-redundant ESTs 总被引:9,自引:1,他引:8
Richard Cooke Monique Raynal Michele Laudi Franoise Grellet Michel Delseny Peter-Christian Morris Danile Guerrier Jrme Giraudat Franoise Quigley Grard Clabault You-Fang Li Rgis Mache Micheline Krivitzky Isabelle Jean-Jacques Gy Martin Kreis Alain Lecharny Yves Parmentier Jacqueline Marbach Jacqueline Fleck Bernadette Clment Gabriel Philipps Christine Herv Claude Bardet Dominique Tremousaygue Bernard Lescure Christophe Lacomme Dominique Roby Marie-Franoise Jourjon Patrick Chabrier Jean-Louis Charpenteau Thierry Desprez Joelle Amselem Helen Chiapello Herman Hfte 《The Plant journal : for cell and molecular biology》1996,9(1):101-124
Nearly 7000 Arabidopsis thaliana -expressed sequence tags (ESTs) from 10 cDNA libraries have been sequenced, of which almost 5000 non-redundant tags have been submitted to the EMBL data bank. The quality of the cDNA libraries used is analysed. Similarity searches in international protein data banks have allowed the detection of significant similarities to a wide range of proteins from many organisms. Alignment with ESTs from the rice systematic sequencing project has allowed the detection of amino acid motifs which are conserved between the two organisms, thus identifying tags to genes encoding highly conserved proteins. These genes are candidates for a common framework in genome mapping projects in different plants. 相似文献
93.
Helen Kennedy 《Brittonia》1995,47(2):156-159
Calathea maasiorum from French Guiana and Surinam is described as new. It belongs toCalathea sectionBreviscapus Bentham. The foliage is patterned with a light green band along the midrib above. This new species has previously been confused
withC. cyclophora Baker from Amazonian Colombia, Brazil, Venezuela, and Guyana but it is distinguished fromC. cyclophora by the absence of bracteoles, the elliptic to obovate leaf blade, and shorter (1–2 cm long) bracts. 相似文献
94.
A Role for Cdk2 Kinase in Negatively Regulating DNA Replication during S Phase of the Cell Cycle 总被引:17,自引:4,他引:13 下载免费PDF全文
Using cell-free extracts made from Xenopus eggs, we show that cdk2-cyclin E and A kinases play an important role in negatively regulating DNA replication. Specifically, we demonstrate that the cdk2 kinase concentration surrounding chromatin in extracts increases 200-fold once the chromatin is assembled into nuclei. Further, we find that if the cdk2–cyclin E or A concentration in egg cytosol is increased 16-fold before the addition of sperm chromatin, the chromatin fails to initiate DNA replication once assembled into nuclei. This demonstrates that cdk2–cyclin E or A can negatively regulate DNA replication. With respect to how this negative regulation occurs, we show that high levels of cdk2–cyclin E do not block the association of the protein complex ORC with sperm chromatin but do prevent association of MCM3, a protein essential for replication. Importantly, we find that MCM3 that is prebound to chromatin does not dissociate when cdk2– cyclin E levels are increased. Taken together our results strongly suggest that during the embryonic cell cycle, the low concentrations of cdk2–cyclin E present in the cytosol after mitosis and before nuclear formation allow proteins essential for potentiating DNA replication to bind to chromatin, and that the high concentration of cdk2–cyclin E within nuclei prevents MCM from reassociating with chromatin after replication. This situation could serve, in part, to limit DNA replication to a single round per cell cycle. 相似文献
95.
Raymond J. Rodgers Tina C. Lavranos Helen F. Rodgers Fiona M. Young Coralie A. Vella 《The Journal of steroid biochemistry and molecular biology》1995,53(1-6):241-246
During folliculogenesis the granulosa cells divide whilst in contact with each other, and so exhibit some of the characteristics of stem cells. In vitro we have shown that bovine granulosa cells from 3–7 mm follicles, like stem cells, divide without the need for a substratum, and produce colonies of cells. Growth factors, bFGF and IGF's, stimulate their division. These cells secrete and assemble a basal lamina, suggesting that the follicular basal lamina is produced by the granulosa cells. They have the morphological characteristics of follicular granulosa cells. Thus this system is ideal for studying the functions of immature granulosa cells because the cells do not spontaneously differentiate or luteinize into luteal cells, as occurs in culture on a substratum. On differentiation into luteal cells in vivo the cells express the steroidogenic enzymes for progesterone production and accumulate β-carotene. During culture of bovine luteal cells we observed that a proportion of the steroidogenic enzyme cholesterol side-chain cleavage cytochrome P450 enzyme became chemically cross-linked to its electron donor, adrenodoxin. P450 enzymes produce oxygen free radicals and oxygen free radicals can cause cross-linking between proteins in close proximity. Cell protect against this damage by the use of antioxidant vitamins. Repleting the cultured luteal cells with β-carotene reduced the amount of cross-linking. We conclude that the high levels of β-carotene in corpora lutea are to protect against damage due to oxygen free radicals generated in the course of progesterone synthesis. 相似文献
96.
Michael J. Barber Andrew S. Zektzer Gerald M. Rosen Helen A. Demos Elmer J. Rauckman 《生物化学与生物物理学报:生物膜》1984,776(1):159-168
Hepatic microsomal membranes, prepared under various conditions that yield either ‘intact’ or ‘disrupted’ microsomal vesicles, have been labeled via the sulfhydryl groups of intrinsic membrane proteins using nitroxide analogs of . Electron paramagnetic resonance spectra revealed the presence of two dominant classes of bound label corresponding to differing degrees of immobilization, the ratio of which were quantitated using a parameter designated the ‘’ ratio. For latent microsomes, the value of this parameter was determined to be and was influenced by factors such as label/protein ratio, incubation period, nitroxide structure, temperature and pH. The ratio was also sensitive to the degree of membrane integrity as revealed by the latency of mannose 6-phosphate activity of glucose-6-phosphohydrolase. In addition, membrane disruption resulted in a corresponding decrease in the order parameter for nitroxide-labeled fatty acids intercalated within the lipid bilayer. The ratio was observed to be dependent upon the method of microsome preparation yielding values of for ‘hypertonically disrupted’ vesicles and for ‘mechanically disrupted’ vesicles. Microsomal marker enzymes such as cytochrome and FAD-containing monooxygenase retained significant levels of functionally following nitroxide incorporation. 相似文献
97.
Huckleberry plants evaluated at 13 sites in the southern Cascade Mountains of Washington State during August 1980 showed significantly lower fruit yields where subjected to heavy ash deposition following the 18 May 1980 eruption of Mt. St. Helens. High insect pollinator mortality is suspected as the major causal factor. The impact of such effects of volcanic activity on Native American subsistence is discussed. 相似文献
98.
99.
A fibrous protein complex extracted fromEscherichia coli B/r by the method of Minkoff and Damadian [2] demonstrates arrowhead complexes when reacted with heavy meromyosin. 相似文献
100.
Helen K. Cooper Teresa Zalewska S. Kawakami K.-A. Hossmann P. Kleihues 《Journal of neurochemistry》1977,28(5):929-934
Abstract— Rats were subjected to cerebral compression ischaemia for 15min and were subsequently recirculated with blood for periods up to 3 h. In vivo incorporation of intravenously administered L-[1–14 C]valine into total brain proteins was found to be severely inhibited (about 20% of controls) after 45 min of recirculation. After 3 h, protein synthesis had increased, the specific radioactivity of proteins then being about 40% of controls. The post-ischaemic inhibition of protein synthesis was accompanied by a breakdown in polyribosomes and a concomitant increase in ribosomal subunits. In vitro incorporation of L-[1–14 C]phenylalanine by a postmitochondrial supernatant system derived from animals subjected to 15 min ischaemia and 15 min recirculation was also severely reduced and showed, in contrast to control animals, no response to the addition of a specific inhibitor of polypeptide chain initiation (Poly(I)). Together with the in vivo accumulation of ribosomal subunits this indicates a block in peptide chain initiation during the early stages of recirculation.
Polyribosomes from animals subjected to 15 min ischaemia without recirculation showed a normal rate of in vitro protein synthesis which was inhibited by Poly(I) to a similar extent as polyribosomes from control animals. These results suggest that the post-ischaemic inhibition in chain initiation develops during the early stages of recirculation rather than during the ischaemic period itself. 相似文献
Polyribosomes from animals subjected to 15 min ischaemia without recirculation showed a normal rate of in vitro protein synthesis which was inhibited by Poly(I) to a similar extent as polyribosomes from control animals. These results suggest that the post-ischaemic inhibition in chain initiation develops during the early stages of recirculation rather than during the ischaemic period itself. 相似文献