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Cyclic ADR-Ribose Synthases (cADPRSs) are one of the NAD-consuming enzymes. It hydrolyzes and cyclizes NAD+ into ADPR and cADPR, which are both Ca2+ channel regulators. Since the activities of cADPRSs involves in multiple biological process, monitoring the activities of these enzymes can provide useful information for studying pathology of the diseases that are closely associated with these enzymes. In this thesis, we have designed and synthesized a series of small molecule probes which can undergo base exchange reaction with the nicotinamide group of NAD+ in the presence of CD38 or activated SARM1. A large red shift of emission wavelength occurred after the base exchange reaction, which provides a powerful tool for detecting activities of this class of enzymes. In the first project, these probes were applied to the detection of the activities of SARM1. Among the 23 probes prepared, PC6 and PC11 showed excellent sensitivity and selectivity in vitro. They are cell-permeant, yet the resulting exchange products are im-permeant, allowing imaging of activated SARM1 in live cells. PC6 has provided the first evidence that SARM1 activation precedes axon degeneration by several hours in live DRG neurons. Moreover, it was also applied in the library screening for SARM1 inhibitor. Dehydronitrosonisodipine (dHNN) was found to has the inhibition ability to SARM1 activation, which is also the first compound ever reported that can inhibit SARM1 activation. PC11 has better fluorescent properties than PC6. With larger absorption and emission wavelength, PC11 provided the first approach for imaging SARM1 activation in vivo. In the second project, we focused on the study of the catalytic mechanism of CD38. Based on the preliminary results of the theoretical studies, we proposed that CD38 catalyzed cyclization and hydrolysis of NAD+ may involve an epoxide intermediate. In this mechanistic study, we have employed our newly developed probe PC6, CD38 mutant and different model compounds. The results of this study strongly supported the hypothesis of an epoxide intermediate in CD38-catalyzed reactions.
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Rapid developments in molecular and systems biology techniques have allowed researchers to unravel many new mechanisms through which plant cells switch over to alternative respiratory pathways. This book is a unique compendium of how and why higher plants evolved alternative respiratory metabolism. It offers a comprehensive review of current research in the biochemistry, physiology, classification and regulation of plant alternative respiratory pathways, from alternative oxidase diversity to functional marker development. The resource provides a broad range of perspectives on the applications of plant respiratory physiology, and suggests brand new areas of research. Other key features: written by an international team of reputed plant physiologists, known for their pioneering contributions to the knowledge of regular and alternative respiratory metabolism in higher plants includes step-by-step protocols for key molecular and imaging techniques advises on regulatory options for managing crop yields, food quality and environment for crop improvement and enhanced food security covers special pathways which are of key relevance in agriculture, particularly in plant post-harvest commodities Primarily for plant physiologists and plant biologists, this authoritative compendium will also be of great value to postdoctoral researchers working on plant respiration, as well as to graduate and postgraduate students and university staff in Plant Science. It is a useful resource for corporate and private firms involved in developing functional markers for breeding programs and controlling respiration for the prevention of post-harvest losses in fruit, vegetables, cut flowers and tubers.
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An understanding of the mechanisms by which plants perceive environmental cues, both physical and chemical, and transduce the signals that influence specific expression of genes, is an area of intensive scientific research. With the completion of the genome sequence of Arabidopsis it is understood now that a larger number of genes encode for proteins involved in signalling cascades and transcription factors. In this volume, different chapters deal with plant receptors, second messengers like calcium ions, phosphoinositides, salicylic acid and nitrous oxide, calcium binding proteins and kinases. In addition to dealing with the response of plants to light, hormones, pathogens, heat, etc. on cellular activity, work currently going on in apoptosis, cell division, and plastid gene expression is also covered in this book.
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