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Repeat proteins are found in almost all cellular systems, where they are involved in diverse molecular recognition processes. Recent studies have suggested that de novo designed repeat proteins may serve as universal binders, and might potentially be used as practical alternative to antibodies. We describe here a novel chemical methodology for producing small libraries of repeat proteins, and screening in parallel the ligand binding of library members. The first stage of this research involved the total synthesis of a consensus-based three-repeat tetratricopeptide (TPR) protein (~14 kDa), via sequential attachment of the respective peptides. Despite the effectiveness of the synthesis and ligation steps, this method was found to be too demanding for the production of proteins containing variable number of repeats. Additionally, the analysis of binding of the individual proteins was time consuming. Therefore, we designed and prepared novel dynamic combinatorial libraries (DCLs), and show that their equilibration can facilitate the formation of TPR proteins containing up to eight repeating units. Interestingly, equilibration of the library building blocks in the presence of the biologically relevant ligands, Hsp90 and Hsp70, induced their oligomerization into forming more of the proteins with large recognition surfaces. We suggest that this work presents a novel simple and rapid tool for the simultaneous screening of protein mixtures with variable binding surfaces, and for identifying new binders for ligands of interest.  相似文献   
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1. Bovine erythrocytes exposed to the action of an enzymic source of hyperoxide radicals (hypoxanthine + xanthine oxidase) exhibited hemolysis, which was prevented by the presence of hyperoxide dismutase. 2. Exposing bovine erythrocyte membranes to the source of hyperoxide radicals resulted in a decrease of (Mg2+ + Na+ + K+)ATPase activity which could be partially prevented by addition of hyperoxide dismutase. 3. The damage observed to erythrocyte membranes under the conditions applied is ascribed to toh formed in the Haber and Weiss reaction since a protection by OH scavengers was also observed.  相似文献   
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The relationships between the apoproteins of intact human serum high density lipoprotein particles, HDL2 and HDL3, have been studied by observing the exchange of radioactively labeled apoproteins between one subclass and the other. This exchange process can be inhibited by chemically crosslinking the apoproteins of either the labeled or unlabeled subclass. These results are consistent with a dynamic relationship between HDL2 and HDL3 which appears dependent upon the association and perhaps the conformation of the apoprotein components of the lipoprotein particles.  相似文献   
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