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211.
Summary The ability of time-averaged restrained molecular dynamics (TARMD) to escape local low-energy conformations and explore conformational space is compared with conventional simulated-annealing methods. Practical suggestions are offered for performing TARMD calculations with ligand-receptor systems, and are illustrated for the complex of the immunosuppressant FK506 bound to Q50R,A95H,K98I triple mutant FKBP-13. The structure of 13C-labeled FK506 bound to triple-mutant FKBP-13 was determined using a set of 87 NOE distance restraints derived from HSQC-NOESY experiments. TARMD was found to be superior to conventional simulated-annealing methods, and produced structures that were conformationally similar to FK506 bound to wild-type FKBP-12. The individual and combined effects of varying the NOE restraint force constant, using an explicit model for the protein binding pocket, and starting the calculations from different ligand conformations were explored in detail.Abbreviations DG
distance geometry
- dmFKBP-12
double-mutant (R42K,H87V) FKBP-12
- FKBP-12
FK506-binding protein (12 kDa)
- FKBP-13
FK506-binding protein (13 kDa)
- HSQC
heteronuclear single-quantum coherence
- KNOE
force constant (penalty) for NOE-derived distance restraints
- MD
molecular dynamics
- NOE
nuclear Overhauser effect
- SA
simulated annealing
- TARMD
molecular dynamics with time-averaged restraints
- tmFKBP-13
triple-mutant (Q50R,A95H,K98I) FKBP-13
- wtFKBP-12
wild-type FKBP-12 相似文献
212.
Pearlman DA 《Journal of biomolecular NMR》1996,8(1):49-66
Summary A new NMR refinement method, FINGAR (FIt NMR using a Genetic AlgoRithm), has been developed, which allows one to determine a weighted set of structures that best fits measured NMR-derived data. This method shows appreciable advantages over commonly used refinement methods. FINGAR generates an ensemble of conformations whose average reproduces the experimental NMR-derived restraints. In addition, a statistical importance weight is assigned to each of the conformations in the ensemble. As a result, one is not limited to simply presenting an envelope of sampled conformers. Instead, one can subsequently focus on a select few conformers of high weight. This is critical, because many structural analyses depend on using discrete conformations, not simply averages or ensembles. The genetic algorithm used by FINGAR allows one to simultaneously and reliably fit against many restraints, and to generate solutions which include as many conformations with non-zero weights as are necessary to generate the best fit. An added benefit of FINGAR is that because the time-consuming step in this method needs only to be performed once, in the beginning of the first run, numerous FINGAR simulations can be performed rapidly. 相似文献
213.
Jan Oscarsson Yoshimitsu Mizunoe Bernt Eric Uhlin David J. Haydon 《Molecular microbiology》1996,20(1):191-199
The Salmonella typhimurium protein SlyAST, originally described as a cytolysin, shows sequence similarities to several known bacterial regulatory proteins. A homologue to the slyASt gene has been localised to min 37 of the Eschericia coli K-12 chromosome and has been designated slyAEC When introduced in trans on a plasmid, the slyAEC gene conferred a haemolytic phenotype on wild-type but not clyA-knockout strains of E. coli K-12. The clyA gene encodes a novel haemolysin that is not expressed by wild-type E. coli under tested laboratory conditions. Western and Northern blot analyses, and DNA-band-shift assays support a model whereby the SlyAEC protein activates clyA expression by binding to the clyA promoter region, thereby supporting the sequence similarity data in suggesting that SlyAST is a haemolysin activator rather than being a haemolysin per se. 相似文献
214.
Two species of diatoms were genetically transformed by introducing plasmid vectors containing the Escherichia coli neomycin phosphotransferase II (npt II ) gene. Expression of the bacterial npt II gene in the diatoms was achieved using the putative promoter and terminator sequences from the acetyl-CoA carboxylase gene from the centric diatom Cyclotella cryptica T13L Reimann, Lewin, and Guillard. The vectors were introduced into C. cryptica and the pennate diatom Navicula saprophila NAVIC1 Lange-Bertalot and Bonik by microprojectile bombardment. Putative transformants were selected based on their ability to grow in the presence of the antibiotic G418, and production of the neomycin phosphotransferase protein by the transformed cells was confirmed by western blotting. The foreign DNA integrated into one or more random sites within the genome of the transformed algal cells, often in the form of tandem repeats. This is the first report of reproducible, stable genetic transformation of a chlorophyll c -containing alga . 相似文献
215.
Gene rearrangements in Chlamydomonas chloroplast DNAs are accounted for by inversions and by the expansion/contraction of the inverted repeat 总被引:1,自引:0,他引:1
To gain insight into the mutational events responsible for the extensive variation of chloroplast DNA (cpDNA) within the green algal genus Chlamydomonas, we have investigated the chloroplast gene organization of Chlamydomonas pitschmannii, a close relative of the interfertile species C. eugametos and C. moewusii whose cpDNAs have been well characterized. At 187 kb, the circular cpDNA of C. pitschmannii is the smallest Chlamydomonas cpDNA yet reported; it is 56 and 105 kb smaller than those of its C. eugametos and C. moewusii counterparts, respectively. Despite this substantial size difference, the arrangement of 77 genes on the C. pitschmannii cpDNA displays only three noticeable differences from the organization of the corresponding genes on the collinear C. eugametos and C. moewusii cpDNAs. These changes in gene order are accounted for by the expansion/contraction of the inverted repeat and one or two inversions in a single-copy region. In land plant cpDNAs, these kinds of events are also responsible for gene rearrangements. The large size difference between the C. pitschmannii and C. eugametos/C. moewusii cpDNAs is mainly attributed to multiple events of deletions/additions as opposed to the usually observed expansion/contraction of the inverted repeat in land plant cpDNAs. We also found that the mitochondrial genome of C. pitschmannii is a circular DNA molecule of 16.5 kb which is 5.5 and 7.5 kb smaller than its C. moewusii and C. eugametos counterparts, respectively. 相似文献
216.
Eric L. Charnov 《Evolutionary ecology》1995,9(3):288-291
Summary Offspring production over the adult lifespan (b/M whereb is the yearly fledgling or offspring production and 1/M is the mean adult lifespan) is an approximate invariant within both birds and mammals. The two taxa differ, however, in that mammals have bothM and b as invariants (b/M = b/M) while birds do not ( is the age at first breeding). Birds have a surprising cancellation in that bothM andb are
–0.25. 相似文献
217.
Specific Activity of Brain Palmitoyl-CoA Pool Provides Rates of Incorporation of Palmitate in Brain Phospholipids in Awake Rats 总被引:4,自引:1,他引:3
Eric Grange Joseph Deutsch Quentin R. Smith Michael Chang Stanley I. Rapoport A. David Purdon 《Journal of neurochemistry》1995,65(5):2290-2298
Abstract: In vivo rates of palmitate incorporation into brain phospholipids were measured in awake rats following programmed intravenous infusion of unesterified [9,10-3 H]palmitate to maintain constant plasma specific activity. Animals were killed after 2–10 min of infusion by microwave irradiation and analyzed for tracer distribution in brain phospholipid and phospholipid precursor, i.e., brain unesterified palmitate and palmitoyl-CoA, pools. [9,10-3 H]Palmitate incorporation into brain phospholipids was linear with time and rapid, with >50% of brain tracer in choline-containing glycerophospholipids at 2 min of infusion. However, tracer specific activity in brain phospholipid precursor pools was low and averaged only 1.6–1.8% of plasma unesterified palmitate specific activity. Correction for brain palmitoyl-CoA specific activity increased the calculated rate of palmitate incorporation into brain phospholipids (0.52 nmol/s/g) by ∼60-fold. The results suggest that palmitate incorporation and turnover in brain phospholipids are far more rapid than generally assumed and that this rapid turnover dilutes tracer specific activity in brain palmitoyl-CoA pool owing to release and recycling of unlabeled fatty acid from phospholipid breakdown. 相似文献
218.
A reevaluation of floral and vegetative morphology together with data from analysis of floral fragrance and flavonoid co-pigment
chemistry suggest that specific status is warranted for the two kinds ofPhalaenopsis violacea Witte known informally as the “Bornean type” and the “Malayan type.”Phalaenopsis violacea var.bellina Rchb.f. is elevated to specific rank, providing a name for the “Bornean type” plants. 相似文献
219.
The effects of weed strips on aphids and aphidophagous predators in an apple orchard 总被引:1,自引:0,他引:1
Eric Wyss 《Entomologia Experimentalis et Applicata》1995,75(1):43-49
Selected weeds were used to attract antagonists of apple aphids in an apple orchard near Berne, Switzerland. In the year before
the experiment, in 1991, the apple aphidsDysaphis plantaginea (Pass.) andAphis pomi (DeGeer) and aphidophagous predators were homogenously distributed in the orchard. In April 1992, weed strips were sown between
tree rows and along the border parallel to the first and the last row of trees in one part (the other part served as control).
In both parts of the orchard, randomly chosen tress were controlled visually in weekly intervals in 1992 and 1993. During
flowering of weeds more aphidophagous predators were observed on the apple trees within the strip-sown area than in the control
area. The most abundant and permanent aphidophagous predators were spiders, predaceous Heteroptera, Coccinellidae, and Chrysopidae.
Both species of aphids were significantly less abundant in the area with weed strips than in the control area during the vegetation
period. The effects of the weed strips on aphidophagous predators, and those of predators on aphids, are discussed. 相似文献
220.
Sixteen new species are proposed in the generaAckermania, Dressleria, Epidendrum, Maxillaria, Oncidium, Rodriguezia, Sigmatostalix, andTrigonidium. All new species are illustrated.Maxillaria vittariifolia L. O. Williams is newly recorded for Peru. A key is provided forTrigonidium of Peru.Trigonidium loretoense Schltr. andT. peruvianum Schltr. are lectotypified. 相似文献