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• W1566585179 abstract "It remains unclear whether polypeptide chains renaturing in vitro from strong denaturants proceed through the same folding pathway as chains released from ribosome within cells. Folding intermediates formed both in vivo and in vitro have been examined using three monoclonal antibodies shown previously to recognize different epitopes of the native P22 tailspike protein (Friguet, B., Djavadi-Ohaniance, L., Haase-Pettingell, C. A., King J., and Goldberg, M. E. (1990) J. Biol. Chem. 265, 10347-10351). The tailspike protein was reconstituted from polypeptide chains unfolded by urea as described by Fuchs et al. (Fuchs, A., Seiderer, C., and Seckler, R. (1991) Biochemistry 30, 6598-6604), and the appearance of immunoreactive forms during the refolding was monitored. The three antibodies discriminated intermediates at different stages in the folding pathway. On the basis of the reconstitution pathway determined from spectroscopic and hydrodynamic measurements by Fuchs et al. (1991), monoclonal antibody (mAb) 236-3 recognized partially folded monomers, mAb 155-3 recognized folded protomers in a protrimer species, and mAb 33-2 recognized the native trimer. The kinetics of appearance of the immunoreactive forms during the in vitro refolding of the protein in crude extracts of phage-infected cells was similar to that observed with the pure tailspike. Thus, the antibodies provided probes for the chain folding and association pathway in vivo. The conformation of the ribosome-bound tailspike polypeptide chains of the infected cells was analyzed with the three antibodies. The antibodies recognizing native trimer and the protrimer did not bind chains associated with the ribosomes. Antibody 236-3, which recognized structured monomers in vitro, bound to the polypeptide chains still associated with ribosomes. This result suggests that steps that take place in solution during in vitro refolding may occur in a ribosome-bound state in vivo." @default.
• W1566585179 created "2016-06-24" @default.
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• W1566585179 date "1994-06-01" @default.
• W1566585179 modified "2023-09-30" @default.
• W1566585179 title "In vitro and ribosome-bound folding intermediates of P22 tailspike protein detected with monoclonal antibodies." @default.
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• W1566585179 doi "https://doi.org/10.1016/s0021-9258(17)40772-1" @default.
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