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Site-directed mutagenesis has been used to produce mutant forms of yeast phosphoglycerate kinase in which the conserved active-site residue, Arg21, has been replaced by a methionine or a lysine. Kinetic results obtained using these mutant enzymes show that their Km for both 3-phospho-D-glycerate and ATP are significantly different from those recorded for the wild-type enzyme. The Vmax for the lysine mutant is reduced by a factor of two from that of the wild-type enzyme whereas the Vmax for the methionine mutant is reduced more than sevenfold. A very clean electron-density-difference map shows little, if any, evidence of a structural change associated with the C-terminal domain, although resonances in the NMR spectra associated with the ATP-binding site (C-terminal domain) are also affected by the mutation as one might expect from the kinetic results. The NMR data show that binding at both the 3-phospho-D-glycerate and the non-productive ATP-binding site (associated with the N-terminal domain) are affected in the mutant in a way which is different to that associated with the wild-type enzyme. These results, taken together with the X-ray and kinetic data, indicate that the non-productive ATP-binding site and the activating anion-binding site are both associated with the basic patch region of yeast phosphoglycerate kinase.  相似文献   
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Aa Gymnastiar (Gym) is a popular Indonesian Muslim preacher who seems to be now at the pinnacle of his fame. He regularly gives advice to the head of state and to ministers and yet at the same time his approach to Islam appeals to all sections of the national Muslim community. His is a familiar face in newspaper columns and above all on television screens; Aa Gym has a masterful command of the media. This article describes and accounts for his popularity and discusses it in terms of continuity and change in the rise and decline of Muslim celebrities in Indonesia. It points out the difference between Gym and some obvious forerunners such as the scholar Hamka, and stresses that the nature of Gym's appeal is new in as much as he does not come from within the circle of traditional families of Muslim ulama . He seems to draw his information as much from secular sources of self-help manuals as from books of Sufi wisdom. Although very popular and influential among the general circle of believers, he is regarded with some suspicion by those who criticize his sufistic leanings and lack of an orthodox Muslim education. The article concludes by arguing that Gym and his approach to the implementation of Muslim precepts is more representative of the nature of Islam in Indonesia today than the activities of terrorists.  相似文献   
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