Heat-treatment method for producing fatty acid-bound alpha-lactalbumin that induces tumor cell death |
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Authors: | Kamijima Tatsuro Ohmura Ayaka Sato Toshiya Akimoto Kaoru Itabashi Miki Mizuguchi Mineyuki Kamiya Masakatsu Kikukawa Takashi Aizawa Tomoyasu Takahashi Masayuki Kawano Keiichi Demura Makoto |
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Institution: | a Division of Life Science, Graduate School of Life Science, Hokkaido University, Kita 10 Nishi 8, Kitaku, Sapporo, Hokkaido 060-0810, Japan b Department of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo 060-0810, Japan c Faculty of Pharmaceutical Sciences, University of Toyama, Toyama 930-0194, Japan d Division of Chemistry, Faculty of Science, Hokkaido University, Sapporo 060-0810, Japan |
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Abstract: | HAMLET (Human Alpha-lactalbumin Made LEthal to Tumor cells), which was identified in human breast milk as an alpha-lactalbumin (LA)-oleic acid complex, kills tumor cells, selectively. Although it may have potential as a therapeutic agent against various tumor cells, only low-volume methods for its production exist. In this study, heat treatment was used to produce complexes from LAs and oleic acid using a simple method. In the case of human LA and oleic acid, heat-treated samples apparently showed much stronger activities than those treated at room temperature, with cytotoxicities equal to that of HAMLET. Furthermore, circular dichroism spectroscopy revealed that heat-treated samples lost their tertiary structure, suggesting a molten globule as oleic acid-bound LA. BLA samples also showed strong activities by heat treatment. Batch production with heat treatment can efficiently convert LAs into tumoricidal complexes. |
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Keywords: | LA alpha-lactalbumin MG molten globule HAMLET human alpha-lactalbumin made lethal to tumor cells CD circular dichroism HLA human alpha-lactalbumin BLA bovine alpha-lactalbumin FCS fetal calf serum PBS phosphate-buffered saline UV ultraviolet |
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