An allosteric ribozyme has been designed using the hammerhead ribozyme as the active site and aflavin-specific RNA aptamer as a regulatory site. We constructed six variants with a series of base pairs in the linker region (stem II). Under single turnover conditions, kinetic studies were carried out in the absence and presence of flavin mononucleotide (FMN). Interestingly, FMN addition did not influence the cleavage rate of constructs with a 5-6 bp linker but stimulated the catalytic activity of those bearing a shorter linker. In particular, the apparent k cat of Rz3 increases by approximately 10-fold upon addition of saturating amounts of FMN. To determine the rate constants( K m4and k cat), the ribozyme regulated most effectively by FMN was further investigated. FMN mainly affected the k cat value, reflecting the rate limiting conformational change step of the overall cleavage reaction, depending on helix formation in stem II. Probably, FMN influences the orientation of structures necessary for the cleavage reaction through stem II formation. The result of chemical modification revealed that binding of FMN to the aptamer domain induced the helix formation in stem II required for catalytic activity. Therefore, a specific FMN-mediated allosteric interaction seems to promote a conformational alteration from an open to a closed structure in stem II. The concept of conformational modification in the allosteric effect is consistent with other allosteric enzymes, suggesting that such a conformational change is a fundamental feature of allosteric enzymes in biological systems.

A mechanical approach to mean field spin modelsWe construct a free field realization of vertex operators of the dilute A L models along with the Felder complex. For Lϭ3, we also study an E 8 structure in terms of the deformed Virasoro currents.

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In the urban-rural fringe of the Bangkok Metropolitan Region, rapid urbanization is creating a land-use mixture of agricultural fields and residential areas. To develop appropriate policies to enhance recycling of municipal solid waste (MSW), current MSW management was investigated in the oboto (local administrative district) of Bang Maenang in Nonthaburi Province, adjoining Bangkok. The authors conducted a structural interview survey with waste-related organizations and local residents, analysed household waste generation, and performed global positioning system (GPS) tracking of municipal garbage trucks. It was found that MSW was collected and treated by local government, private-sector entities, and the local community separately. Lack of integrated management of these entities complicated waste flow in the study area, and some residences were not served by MSW collection. Organic waste, such as kitchen garbage and yard waste, accounted for a large proportion of waste generation but was underutilized. Through GPS/GIS analysis, the waste collection rate of the generated waste amount was estimated to be 45.5- 51.1% of total generation.