caa a22 4500 475743989 CHVBK 20180406123507.0 cr unu---uuuuu 170329e20000801xx s 000 0 eng 10.1023/A:1026593113633 doi (NATIONALLICENCE)springer-10.1023/A:1026593113633 Chaperone-Mediated Refolding of Recombinant Prochymosin [Elektronische Daten] [Chongjuan Wei, Yuying Zhang, Kaiyu Yang] It has been verified that prochymosin is characterized by a two-stage refolding: dilution of unfolded protein into pH 11 buffer followed by neutralization at pH 8; the high-pH step is indispensable. Here we demonstrate that one-stage refolding around pH 8 can be achieved when GroE or 10-fold molar excess (rather than catalytic concentration) of protein disulfide isomerase (PDI) over prochymosin is present. The helping effect varies with the oxidation states of prochymosin. GroE and PDI increase the reactivation of the unfolded, partially reduced and the unfolded, oxidized prochymosin from 5% to 40% and from 50% to 100%, respectively. For the unfolded and fully reduced prochymosin, GroE does not have a positive effect, whereas PDI promotes renaturation from 2% to 28%. Based on our previous and present observations, we propose that at pH 8 there may be two kinds of incorrect interactions within and between prochymosin polypeptides leading to unproductive pathways: one prevents disulfide rearrangement, which can be avoided by high pH; the other interferes with acquisition of native conformation, which can be relieved by GroE and PDI. Plenum Publishing Corporation, 2000 GroEL nationallicence GroES nationallicence protein disulfide isomerase nationallicence prochymosin nationallicence refolding nationallicence Wei Chongjuan Institute of Microbiology, Chinese Academy of Sciences, 100080, Beijing, China aut Zhang Yuying Institute of Microbiology, Chinese Academy of Sciences, 100080, Beijing, China aut Yang Kaiyu Institute of Microbiology, Chinese Academy of Sciences, 100080, Beijing, China aut Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/6(2000-08-01), 449-456 0277-8033 19:6<449 2000 19 10930 https://doi.org/10.1023/A:1026593113633 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1026593113633 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wei Chongjuan Institute of Microbiology, Chinese Academy of Sciences, 100080, Beijing, China aut NATIONALLICENCE 700 1- Zhang Yuying Institute of Microbiology, Chinese Academy of Sciences, 100080, Beijing, China aut NATIONALLICENCE 700 1- Yang Kaiyu Institute of Microbiology, Chinese Academy of Sciences, 100080, Beijing, China aut NATIONALLICENCE 773 0- Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/6(2000-08-01), 449-456 0277-8033 19:6<449 2000 19 10930 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer