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21.
A. E. Solovchenko M. N. Merzlyak O. B. Chivkunova I. V. Reshetnikova I. Khozina-Goldberg S. Didi-Cohen Z. Cohen 《Moscow University Biological Sciences Bulletin》2008,63(1):44-48
Parietochloris incisa is a unicellular freshwater green alga capable of accumulating high amounts of the valuable long-chain polyunsaturated arachidonic acid (AA) in triacylglycerols (TAG) of cytoplasmic oil bodies. To find the cultivation conditions providing maximum AA yield, the effects of illumination and N-availability on the dry weight (DW), chlorophyll, carotenoid, and AA content were studied. Under nitrogen starvation, TAG accounted for over 30% of dry weight (DW) and the AA content became as high as about 55% of total fatty acids. For biomass accumulation, light intensity of ca 400 μE m?2 s?1 was found to be optimal for growing P. incisa on a complete medium. Lower light intensities (or a higher cell density of inoculum) resulted in a higher AA yield when the alga was cultivated on nitrogen-free media. In the absence of nitrogen, algal cells were unable to cope with high illumination and suffered from photooxidative damage, whereas the nutrientsufficient culture survived under such illumination conditions, probably due to accumulation of carotenoids. Nitrogen-deprived P. incisa cells displayed elevated sensitivity to light. 相似文献
22.
Met and EGF receptors can induce a decrease in intercellular adhesion and an increase in cell motility, which is a cause of metastatic progressions. Therefore, mechanisms of interaction in receptor tyrosine kinase and proteins of intercellular contacts attract the attention of researchers. The main protein that provides cellular adhesion is E-cadherin. Earlier, we have shown that the intracellular Met localization was dependent on function of E-cadherin. In the present work, we have found that localization of the EGF receptor also was determined by adhesion stability. Loss of intercellular contacts in HBL-100 cells leads to the EGF receptor being not stabilized at the cell membrane. A comparative study of MAP kinase activation by growth factors was carried out in cells differing by their intercellular adhesion states. It has been established that E-cadherin is able to modulate level and duration of activation of ERK kinase. The presented results allow for the suggestion to be made that not only intracellular localization, but also the intracellular signal pathway activated by Met and EGF receptors, depend on the E-cadherin function, which in turn can determine the specificity of cellular response. 相似文献
23.
Two temperature-sensitive mutants--AP 16 and AP 18 were isolated after the treatment of E. coli AB2500 strain with two mutagens (acridine orange and 5-bromuracil). The mutants obtained proved to be sensitive and formed revertants when treated with the following agents: N-methyl-N'-nitro-N-nitrosoguanidine, hydroxylamine, nitrous acid, sodium metabisulfite, methylmethansulfonate, and proflavine. Introduction into the mentioned strains of additional mutation causing elevation of their sensitivity to crystal violet increased somewhat their capacity to form revertants under the effect of proflavine and methylmethansulfonate. 相似文献
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Shikimate kinase (SK) and other enzymes in the shikimate pathway are potential targets for developing non-toxic antimicrobial agents, herbicides, and anti-parasite drugs, because the pathway is essential in the above species but is absent from mammals. The crystal structure of Mycobacterium tuberculosis SK (MtSK) in complex with MgADP has been determined at 1.8 A resolution, revealing critical information for the structure-based design of novel anti-M. tuberculosis agents. MtSK, with a five-stranded parallel beta-sheet flanked by eight alpha-helices, has three domains: the CORE domain, the shikimate-binding domain (SB), and the LID domain. The ADP molecule is bound with its adenine moiety sandwiched between the side-chains of Arg110 and Pro155, its beta-phosphate group in the P-loop, and the alpha and beta-phosphate groups hydrogen bonded to the guanidinium group of Arg117. Arg117 is located in the LID domain, is strictly conserved in SK sequences, is observed for the first time to interact with any bound nucleotide, and appears to be important in both substrate binding and catalysis. The crystal structure of MtSK (this work) and that of Erwinia chrysanthemi SK suggest a concerted conformational change of the LID and SB domains upon nucleotide binding. 相似文献
27.
The crystal structures of Thermus thermophilus phenylalanyl-tRNA synthetase (PheRS) complexed with phenylalanine and phenylalaninyl-adenylate (PheOH-AMP), the synthetic analogue of phenylalanyl-adenylate, have been determined at 2.7A and 2.5A resolution, respectively. Both Phe and PheOH-AMP are engulfed in the active site cleft of the catalytic alpha-subunit of PheRS, and neither makes contact with the PheRS beta-subunit. The conformations and binding of Phe are almost identical in both complexes. The recognition of Phe by PheRS is achieved through a mixture of multiple van der Waals interactions and hydrogen bonds. The side-chain of the Phe substrate is sandwiched between the hydrophobic side-chains of Phealpha258 and Phealpha260 on one side, and the main-chain atoms of the two adjacent beta-strands on the other. The side-chains of Valalpha261 and Alaalpha314 form the back wall of the amino acid binding pocket. In addition, PheRS residues (Trpalpha149, Seralpha180, Hisalpha178, Argalpha204, Glnalpha218, and Glualpha220) form a total of seven hydrogen bonds with the main-chain atoms of Phe. The conformation of PheOH-AMP and the network of interactions of its AMP moiety with PheRS are reminiscent of the other class II synthetases. The structural similarity between PheRS and histidyl-tRNA synthetase extends to the amino acid binding site, which is normally unique for each enzyme. The complex structures suggest that the PheRS beta-subunit may affect the first step of the reaction (formation of phenylalanyl-adenylate) through the metal-mediated conserved alpha/beta-subunit interface. The modeling of tyrosine in the active site of PheRS revealed no apparent close contacts between tyrosine and the PheRS residues. This result implies that the proofreading mechanism against activated tyrosine, rather than direct recognition, may play the major role in the PheRS specificity. 相似文献
28.
L S Reshetnikova C O Reiser N K Schirmer H Berchtold R Storm R Hilgenfeld M Sprinzl 《Journal of molecular biology》1991,221(2):375-377
The intact elongation factor Tu from the extreme thermophile Thermus thermophilus has been crystallized as a complex with the GTP analogue guanosine-5'-(beta,gamma-imido)triphosphate. The crystals are very stable in the X-ray beam and diffract to 1.9 A resolution. They exhibit space group C2, with a = 150.3(6) A, b = 99.6(3) A, c = 40.1(1) A, beta = 95.4(2) degrees, and contain one elongation factor Tu molecule per asymmetric unit. 相似文献
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30.
Kumar VS O'Neall-Hennessey E Reshetnikova L Brown JH Robinson H Szent-Györgyi AG Cohen C 《Biophysical journal》2011,101(9):2185-2189
We have determined the crystal structure of a phosphorylated smooth-muscle myosin light chain domain (LCD). This reconstituted LCD is of a sea scallop catch muscle myosin with its phosphorylatable regulatory light chain (RLC SmoA). In the crystal structure, Arg16, an arginine residue that is present in this isoform but not in vertebrate smooth-muscle RLC, stabilizes the phosphorylation site. This arginine interacts with the carbonyl group of the phosphorylation-site serine in the unphosphorylated LCD (determined previously), and with the phosphate group when the serine is phosphorylated. However, the overall conformation of the LCD is essentially unchanged upon phosphorylation. This result provides additional evidence that phosphorylation of the RLC is unlikely to act as an on-switch in regulation of scallop catch muscle myosin. 相似文献