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71.
Frueh DP Arthanari H Koglin A Vosburg DA Bennett AE Walsh CT Wagner G 《Nature》2008,454(7206):903-906
Non-ribosomal peptide synthetases (NRPS) and polyketide synthases (PKS) produce numerous secondary metabolites with various therapeutic/antibiotic properties. Like fatty acid synthases (FAS), these enzymes are organized in modular assembly lines in which each module, made of conserved domains, incorporates a given monomer unit into the growing chain. Knowledge about domain or module interactions may enable reengineering of this assembly line enzymatic organization and open avenues for the design of new bioactive compounds with improved therapeutic properties. So far, little structural information has been available on how the domains interact and communicate. This may be because of inherent interdomain mobility hindering crystallization, or because crystallized molecules may not represent the active domain orientations. In solution, the large size and internal dynamics of multidomain fragments (>35 kilodaltons) make structure determination by nuclear magnetic resonance a challenge and require advanced technologies. Here we present the solution structure of the apo-thiolation-thioesterase (T-TE) di-domain fragment of the Escherichia coli enterobactin synthetase EntF NRPS subunit. In the holoenzyme, the T domain carries the growing chain tethered to a 4'-phosphopantetheine whereas the TE domain catalyses hydrolysis and cyclization of the iron chelator enterobactin. The T-TE di-domain forms a compact but dynamic structure with a well-defined domain interface; the two active sites are at a suitable distance for substrate transfer from T to TE. We observe extensive interdomain and intradomain motions for well-defined regions and show that these are modulated by interactions with proteins that participate in the biosynthesis. The T-TE interaction described here provides a model for NRPS, PKS and FAS function in general as T-TE-like di-domains typically catalyse the last step in numerous assembly-line chain-termination machineries. 相似文献
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Bond J Roberts E Springell K Lizarraga SB Lizarraga S Scott S Higgins J Hampshire DJ Morrison EE Leal GF Silva EO Costa SM Baralle D Raponi M Karbani G Rashid Y Jafri H Bennett C Corry P Walsh CA Woods CG 《Nature genetics》2005,37(4):353-355
Autosomal recessive primary microcephaly is a potential model in which to research genes involved in human brain growth. We show that two forms of the disorder result from homozygous mutations in the genes CDK5RAP2 and CENPJ. We found neuroepithelial expression of the genes during prenatal neurogenesis and protein localization to the spindle poles of mitotic cells, suggesting that a centrosomal mechanism controls neuron number in the developing mammalian brain. 相似文献
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Nitrogen is a key element to control the growth and yield of crops. Fertilizer urea nitrogen (N) 60,45, and 30 kg/hm2 was applied at three different stages, midtillering, panicle initiation, and flowering, of the growth and development of
rice plants, respectively. At both midtillering and panicle initiation, the total activity of glutamine synthetase (GS) in
rice roots and leaves was incrased remarkably as a result of a large amount of ammonia absorbed by roots. Native-PAGE and
activity staining showed that the increase of total activity in rice roots and leaves was due to the synthesis of GSrb in
roots and GS2 in leaves and that the activity of GSra in roots and GS1 in leaves remained constant. The results showed that
the assimilation of external nitrogen was carried out by GSrb but not GSra in rice roots and that the activitry of GS2 was
induced also by the external nitrogen, and that GSrb played main role in meeting the needs of the rapid tillering for nitrogen.
At flowering, the activity of GS in rice roots and leaves did not change almost after topdressing. These rssults suggest that
the change of GS activity in rice roots may use as a measure of the utilization efficiency of the fertilizer.
Supported by the National Natural Science Foundation of China, Natural Science Foundation of Hubei Province, and International
Rice Institute, P. O. Box 933 1099 Manila, Philippines
Zhang Chufu: born in 1946, Professor, to whom Correspondence should be addressed 相似文献
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