| Автор | HIRAYAMA, Atsushi |
| Автор | FUJIO, Hajime |
| Автор | DOHI, Yoshitane |
| Автор | TAKAGAKI, Yutaka |
| Автор | KONDO, Kiyoshi |
| Автор | AMANO, Tsunehisa |
| Дата выпуска | 1981 |
| dc.description | Two derivatives of hen egg-white lysozyme (lysozyme) with single substitutions of a 2,4-dinitrophenyl (DNP) residue were prepared. The reaction of lysozyme with a 10-fold molar excess of 2,4-dinitrobenzene sulfonic acid provided one mono-DNP substituted lysozyme (DNP<sub>1</sub>-33lysozyme), which was purified by ion-exchange chromatographies. The other one (DNP<sub>1</sub>-96lysozyme) was prepared as follows. After maleylation of lysozyme in the presence of a 7-fold molar excess of maleic anhydride, the derivative with one free amino group was purified on DE-52. This material was dinitrophenylated with 2,4-dinitrobenzene sulfonic acid and the mono-DNP substituted derivative was purified on DE-52. DNP<sub>1</sub>-96lysozyme was finally purified on SE-Sephadex C-25, after demaleylation at pH 3.5, at 37°C, for 5 days. DNP<sub>1</sub>-33lysozyme and DNP<sub>1</sub>-96lysozyme each migrated as a single band with slower mobility than that of native lysozyme. On reduction, carboxymethylation and chymotrypsin digestion, both mono-DNP substituted lysozymes yielded a single yellow peptide. The amino acid compositions or partial sequence of these peptides indicated that lysine-33 and lysine-96 were the only dinitrophenylated residues in DNP<sub>1</sub>-33lysozyme and DNP<sub>1</sub>-96lysozyme, respectively. DNP<sub>1</sub>-33lysozyme and DNP<sub>1</sub>-96lysozyme showed antigenic reactivities equal to that of native lysozyme with antisera to lysozyme. The DNP residues on the protein were accessible to anti-DNP antibodies, but the affinities of DNP<sub>1</sub>-33lysozyme to anti-DNP antibodies were lower than those of DNP<sub>1</sub>-96lysozyme. This result is discussed with respect to the local environments of the DNP residues in these proteins. |
| Формат | application.pdf |
| Издатель | Oxford University Press |
| Копирайт | © 1981, by the Japanese Biochemical Society |
| Тема | Articles |
| Название | Preparations and Immunological Characterizations of Two Lysozyme Derivatives Dinitrophenylated at Lysine-33 and Lysine-96, Respectively |
| Тип | research-article |
| Electronic ISSN | 1756-2651 |
| Print ISSN | 0021-924X |
| Журнал | Journal of Biochemistry |
| Том | 89 |
| Первая страница | 963 |
| Последняя страница | 974 |
| Аффилиация | HIRAYAMA Atsushi; Department of Immunology, Research Institute for Microbial Diseases |
| Аффилиация | FUJIO Hajime; Department of Immunology, Research Institute for Microbial Diseases |
| Аффилиация | DOHI Yoshitane; Department of Bacteriology, Osaka University Medical School |
| Аффилиация | TAKAGAKI Yutaka; Department of Immunology, Research Institute for Microbial Diseases |
| Аффилиация | KONDO Kiyoshi; Department of Immunology, Research Institute for Microbial Diseases |
| Аффилиация | AMANO Tsunehisa; Department of Immunology, Research Institute for Microbial Diseases |
| Выпуск | 3 |
