Halophilic Properties of Metal Binding Protein Characterized by High Histidine Content from Chromohalobacter salexigens DSM3043 |
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Authors: | Rui Yamaguchi Tsutomu Arakawa Hiroko Tokunaga Matsujiro Ishibashi Masao Tokunaga |
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Institution: | (1) Biochemistry and Applied Biosciences, The United Graduate School of Agricultural Sciences, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan;(2) Alliance Protein Laboratory, 3957 Corte Cancion, Thousand Oaks, CA 91360, USA;(3) Applied and Molecular Microbiology, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan; |
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Abstract: | Periplasmic metal binding protein characterized by high histidine content was cloned from moderate halophile, Chromohalobacter salexigens. The protein, termed histidine-rich metal binding protein (HP), was expressed in and purified from E. coli as a native form. HP bound to Ni- and Cu-loaded chelate columns with high affinity, and Co- and Zn-columns with moderate
affinity. Although the secondary structure was not grossly altered by the addition of 0.2–2.0 M NaCl, the thermal transition
pattern was considerably shifted to higher temperature with increasing salt concentration: melting temperature was raised
by ~20 °C at 2.0 M NaCl over the melting temperature at 0.2 M NaCl. HP showed reversible refolding from thermal melting in
0.2–1.15 M NaCl, while it formed irreversible aggregates upon thermal melting at 2 M NaCl. Addition of 0.01–0.1 mM NiSO4 stabilized HP against thermal melting with high reversibility, while addition above 0.5 mM resulted in irreversible melting
due to aggregation. |
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