caa a22 4500 397493967 CHVBK 20180308164516.0 cr unu---uuuuu 161202e199502 xx s 000 0 eng 10.1093/jb/117.2.384 doi (NATIONALLICENCE)oxford-10.1093/jb/117.2.384 The Folding Characteristics of Tryptophanase from Escherichia coli [Elektronische Daten] [Tomohiro Mizobata, Yasushi Kawata] The unfolding and refolding characteristics of Escherichia coli tryptophanase (tryptophan indole-lyase) [EC 4.1.99.1] in guanidine hydrochloride were studied. Tryptophanase unfolded by first dissociating its coenzyme, pyridoxal 5'-phosphate, from the active site. This dissociation caused a significant destabilization of structure, and global unfolding of the protein followed. During this global unfolding step, an intermediate was formed which had a strong tendency to aggregate irreversibly, as detected by light scattering experi ments. Tryptophanase was unable to refold quantitatively after unfolding in 4 M guanidine hydrochloride. The low refolding yield was due to non-specific aggregation which occurs during refolding. Various conditions which limited this aggregation were probed, and it was found that by initiating the refolding reaction at low temperature, the aggregation of tryptophanase folding intermediates during the reaction could be avoided to a certain extent, and the refolding yield improved. © BY THE JOURNAL OF BIOCHEMISTRY Regular Papers nationallicence aggregation nationallicence folding nationallicence oligomeric protein nationallicence tryptophanase nationallicence Mizobata Tomohiro Department of Chemistry, Faculty of Science, Kyoto University Sakyo-ku, Kyoto 606-01 aut Kawata Yasushi Department of Chemistry, Faculty of Science, Kyoto University Sakyo-ku, Kyoto 606-01 aut The Journal of Biochemistry Oxford University Press 117/2(1995-02), 384-391 0021-924X 117:2<384 1995 117 jbchem https://doi.org/10.1093/jb/117.2.384 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/jb/117.2.384 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Mizobata Tomohiro Department of Chemistry, Faculty of Science, Kyoto University Sakyo-ku, Kyoto 606-01 aut NATIONALLICENCE 700 1- Kawata Yasushi Department of Chemistry, Faculty of Science, Kyoto University Sakyo-ku, Kyoto 606-01 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 117/2(1995-02), 384-391 0021-924X 117:2<384 1995 117 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford