Discovery of the ammonium substrate site on glutamine synthetase, a third cation binding site

Glutamine synthetase (GS) catalyzes the ATP-dependent condensation of ammonia and glutamate to yield glutamine, ADP, and inorganic phosphate in the presence of divalent cations. Bacterial GS is an enzyme of 12 identical subunits, arranged in two rings of 6, with the active site between each pair of subunits in a ring. In earlier work, we have reported the locations within the funnel-shaped active site of the substrates glutamate and ATP and of the two divalent cations, but the site for ammonia (or ammonium) has remained elusive. Here we report the discovery by X-ray crystallography of a binding site on GS for monovalent cations, Tl+ and Cs+, which is probably the binding site for the substrate ammonium ion. Fourier difference maps show the following. (1) Tl+ and Cs+ bind at essentially the same site, with ligands being Glu 212, Tyr 179, Asp 50', Ser 53' of the adjacent subunit, and the substrate glutamate. From its position adjacent to the substrate glutamate and the cofactor ADP, we propose that this monovalent cation site is the substrate ammonium ion binding site. This proposal is supported by enzyme kinetics. Our kinetic measurements show that Tl+, Cs+, and NH4+ are competitive inhibitors to NH2OH in the gamma-glutamyl transfer reaction. (2) GS is a trimetallic enzyme containing two divalent cation sites (n1, n2) and one monovalent cation site per subunit. These three closely spaced ions are all at the active site: the distance between n1 and n2 is 6 A, between n1 and Tl+ is 4 A, and between n2 and Tl+ is 7 A. Glu 212 and the substrate glutamate are bridging ligands for the n1 ion and Tl+. (3) The presence of a monovalent cation in this site may enhance the structural stability of GS, because of its effect of balancing the negative charges of the substrate glutamate and its ligands and because of strengthening the "side-to-side" intersubunit interaction through the cation-protein bonding. (4) The presence of the cofactor ADP increases the Tl+ binding to GS because ADP binding induces movement of Asp 50' toward this monovalent cation site, essentially forming the site. This observation supports a two-step mechanism with ordered substrate binding: ATP first binds to GS, then Glu binds and attacks ATP to form gamma-glutamyl phosphate and ADP, which complete the ammonium binding site. The third substrate, an ammonium ion, then binds to GS, and then loses a proton to form the more active species ammonia, which attacks the gamma-glutamyl phosphate to yield Gln. (5) Because the products (Glu or Gln) of the reactions catalyzed by GS are determined by the molecule (water or ammonium) attacking the intermediate gamma-glutamyl phosphate, this negatively charged ammonium binding pocket has been designed naturally for high affinity of ammonium to GS, permitting glutamine synthesis to proceed in aqueous solution.     

用于烯烃复分解反应的钌茚基催化剂的合成研究进展

基于钌金属的烯烃复分解反应现今已成为一种至关重要且快速有效的催化反应类型,在聚合领域以及现代有机合成的应用方面起着改革性的推进作用。作为烯烃复分解反应催化剂的一种,钌茚基催化剂在该领域得到成功应用。结合近年来钌茚基催化剂在烯烃复分解反应中的合成研究进展,简述了其发展过程,介绍了该领域创新性的研究成果,重点论述了一些新型钌茚基催化剂的合成方法,并且探讨了辅助配体的结构对催化剂稳定性和活性的影响,最后对其发展方向以及应用前景进行了展望。     

Erratum to: Charge transfer in graphene/polymer interfaces for CO2 detection

Nano Research - The order of the authors in the original version of this article was unfortunately incorrect on the first page and the first page of the ESM. Instead of Myungwoo Son1, Yusin Pak1,...     

Localization of atrial natriuretic peptide receptors in the rat tongue and hard palate

The effect of a naturally occurring plant phenolic constituent (the acylphloroglucinol derivative, jensenone, derived from Eucalyptus jensenii) on the food intake of two folivorous marsupials, the common ringtail (Pseudocheirus peregrinus) and the common brushtail possum (Trichosurus vulpecula) was studied. When fed diets containing varying concentrations of jensenone, both species regulated their intake of jensenone so as not to exceed a ceiling intake. This ceiling was about twice as high for common ringtails as for common brushtails from northern Australia. Southern populations of common ringtails showed greatly reduced capacities to tolerate jensenone. When common brushtails were injected (0.5 mg.kg-0.75 body mass) with ondansetron (a selective antagonist of serotonin 5HT3 receptors), they ate significantly more jensenone than animals injected with physiological saline. The same pattern was observed when common ringtails were fed diets containing both jensenone and ondansetron (0.0035 mg.g-1 wet mass of diet). Ondansetron injection had no effect on food intake when the food did not contain jensenone while the addition of higher doses of ondansetron to diets of common ringtails very slightly reduced food intakes of a non-jensenone diet. When common brushtails were given 50 mg of jensenone by gastric lavage, their average subsequent intake of dietary jensenone matched the difference between the daily threshold and the dose given, although the response of individuals was highly variable. Lavage with water alone had no effect on subsequent jensenone intake compared with the pre-dose period. We interpret these results as evidence that the antifeedant effects of jensenone and related compounds are partly mediated by serotonin action on 5HT3 receptors most likely via "nausea" to condition a food aversion.     

Periodic orbits: a new language for neuronal dynamics

A new nonlinear dynamical analysis is applied to complex behavior from neuronal systems. The conceptual foundation of this analysis is the abstraction of observed neuronal activities into a dynamical landscape characterized by a hierarchy of "unstable periodic orbits" (UPOs). UPOs are rigorously identified in data sets representative of three different levels of organization in mammalian brain. An analysis based on UPOs affords a novel alternative method of decoding, predicting, and controlling these neuronal systems.     

Infective endocarditis complicated with rapidly progressive glomerulonephritis: a case report

Despite a general national decline in criminal activities in the 1990s, juvenile criminal offenses continue to increase, (including violent, property, and delinquency acts). In addition increased numbers of children are being held in juvenile jails. It is all but impossible for pediatric health providers to think that "their patients" and "their practices" are immune from the epidemic of crime that affects and is caused by "just kids."     

Rerouting of the intratemporal facial nerve: an analysis of the literature

Anterior rerouting of the intratemporal facial nerve in the infratemporal fossa approach is employed to access to the jugular bulb, hypotympanum, and lateral skull base, whereas posterior rerouting of the facial nerve, as employed in the transcochlear craniotomy, is most frequently used for surgery of the posterior fossa, cerebellopontine angle, prepontine region, and petrous apex. Facial nerve rerouting may lead to facial paresis or paralysis. This review of the literature is intended to define the physiologic "cost" of these procedures, so that the neurotologic surgeon can determine if the morbidity incurred in these techniques is worth the resultant exposure. Inconsistencies in reporting facial function places into question the validity of some of the cumulative data reported. Postoperatively, grades I-II facial nerve function was seen in 91% of patients undergoing short anterior rerouting, 74% of patients undergoing long anterior rerouting, and 26% of patients undergoing posterior complete rerouting. Although facial nerve rerouting allows unhindered exposure to previously inaccessible regions, it is achieved at the cost of facial nerve function. Facial nerve dysfunction increases with the length of facial nerve rerouted.     

Modelling co-operation in the design of knowledge production systems: The Made In'Coop method

This paper presents the latest developments of the MadeIn 'Coop method for modelling the human-machine and human-human co-operation process, and an application of this method for the design of a more co-operative version of the C³I System CHEOPS. We first consider that the design of software systems for organizations is tied more and more to the perspective of compound Knowledge Production Systems that link humans and machines engaged in a co-operative problem solving process. After exposing the four principles upon which MadeIn 'Coop rests for modelling co-operation, we present an artificial problem solving dialogue between CHEOPS and its users. Consistent with the Group Cognitive Processes Theory framework, we propose a dialogue analysis according to two complimentary points of view: the Collective Problem Solving model, and the Coordination model. This analysis should help system designers to identify new system functionalities to assist problem solving.(C³I) Command Control Communication Intelligence Systems     

A 1-Local Asymptotic 13/9-Competitive Algorithm for Multicoloring Hexagonal Graphs

In the frequency allocation problem, we are given a mobile telephone network, whose geographical coverage area is divided into cells, wherein phone calls are serviced by assigning frequencies to them so that no two calls emanating from the same or neighboring cells are assigned the same frequency. The problem is to use the frequencies efficiently, i.e., minimize the span of frequencies used. The frequency allocation problem can be regarded as a multicoloring problem on a weighted hexagonal graph. In this paper, we give a 1-local asymptotic 4/3-competitive distributed algorithm for multicoloring a triangle-free hexagonal graph, which is a special case of hexagonal graph. Based on this result, we then propose a 1-local asymptotic13/9-competitive algorithm for multicoloring the (general-case) hexagonal graph, thereby improving the previous 1-local 3/2-competitive algorithm. A preliminary version of this paper appeared in the Proceedings of the 13th Annual International Computing and Combinatorics Conference (COCOON 2007), LNCS 4598, pp. 526–536. Y. Zhang research was supported by European Regional Development Fund (ERDF). F.Y.L. Chin research was supported by Hong Kong RGC Grant HKU-7113/07E. H. Zhu research was supported by National Natural Science Fund (grant #60496321).