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Walter TS Meier C Assenberg R Au KF Ren J Verma A Nettleship JE Owens RJ Stuart DI Grimes JM 《Structure (London, England : 1993)》2006,14(11):1617-1622
Crystallization remains a critical step in X-ray structure determination. Because it is not generally possible to rationally predict crystallization conditions, commercial screens have been developed which sample a wide range of crystallization space. While this approach has proved successful in many cases, a significant number of proteins fail to crystallize despite being soluble and monodispersed. It is established that chemical modification can facilitate the crystallization of otherwise intractable proteins. Here we describe a method for the reductive methylation of lysine residues which is simple, inexpensive, and efficient, and report on its application to ten proteins. We describe the effect of methylation on the physico-chemical properties of these proteins, and show that it led to diffraction-quality crystals from four proteins and structures for three that had hitherto proved refractory to crystallization. The method is suited to both low- and high-throughput laboratories. 相似文献
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Sutton G Fry E Carter L Sainsbury S Walter T Nettleship J Berrow N Owens R Gilbert R Davidson A Siddell S Poon LL Diprose J Alderton D Walsh M Grimes JM Stuart DI 《Structure (London, England : 1993)》2004,12(2):341-353
As part of a high-throughput structural analysis of SARS-coronavirus (SARS-CoV) proteins, we have solved the structure of the non-structural protein 9 (nsp9). This protein, encoded by ORF1a, has no designated function but is most likely involved with viral RNA synthesis. The protein comprises a single beta-barrel with a fold previously unseen in single domain proteins. The fold superficially resembles an OB-fold with a C-terminal extension and is related to both of the two subdomains of the SARS-CoV 3C-like protease (which belongs to the serine protease superfamily). nsp9 has, presumably, evolved from a protease. The crystal structure suggests that the protein is dimeric. This is confirmed by analytical ultracentrifugation and dynamic light scattering. We show that nsp9 binds RNA and interacts with nsp8, activities that may be essential for its function(s). 相似文献
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Sarah Sainsbury Jingshan Ren Joanne E Nettleship Nigel J Saunders David I Stuart Raymond J Owens 《BMC structural biology》2010,10(1):10
Background
Survival of the human pathogen, Neisseria meningitidis, requires an effective response to oxidative stress resulting from the release of hydrogen peroxide by cells of the human immune system. In N. meningitidis, expression of catalase, which is responsible for detoxifying hydrogen peroxide, is controlled by OxyR, a redox responsive LysR-type regulator. OxyR responds directly to intracellular hydrogen peroxide through the reversible formation of a disulphide bond between C199 and C208 in the regulatory domain of the protein. 相似文献19.
Jingshan?Ren Sarah?Sainsbury Nick?S?Berrow David?Alderton Joanne?E?Nettleship David?K?Stammers Nigel?J?Saunders Raymond?J?OwensEmail author 《BMC structural biology》2005,5(1):13
Background
The NMB0736 gene of Neisseria meningitidis serogroup B strain MC58 encodes the putative nitrogen regulatory protein, IIANtr (abbreviated to NM-IIANtr). The homologous protein present in Escherichia coli is implicated in the control of nitrogen assimilation. As part of a structural proteomics approach to the study of pathogenic Neisseria spp., we have selected this protein for structure determination by X-ray crystallography. 相似文献20.
Forced extra-pair copulations (FEPC) are frequent and mate guarding well developed among common murres. Male murres were at the colony almost continuously through the pre-laying period, but females were present only infrequently, and the frequency of FEPCs was significantly correlated with the relative number of males present. Males vigorously defended their partners from other males and females attempted to resist extra-pair matings. Females whose mates were absent were particularly vulnerable to FEPCs. The number of extra-pair copulations performed by males was estimated to vary between 0–32/season. The reproductive successes of males not performing FEPC, and those performing at the mean and maximum rate are estimated. Males most active in FEPC may substantially increase their reproductive success compared with males not performing FEPC. Ecological conditions in the common murre probably favour prolonged sperm storage, which in turn provides opportunities for sperm competition and favours both effective mate guarding and extra-pair copulations. 相似文献