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A Neutron crystallographic analysis of a cubic porcine insulin at pD 6.6

pD6.6におけるインスリン正方晶の中性子結晶構造解析

石川 卓哉*; 茶竹 俊行*; 大西 裕季*; 田中 伊知朗*; 栗原 和男; 黒木 良太; 新村 信雄*

Ishikawa, Takuya*; Chatake, Toshiyuki*; Onishi, Yuki*; Tanaka, Ichiro*; Kurihara, Kazuo; Kuroki, Ryota; Niimura, Nobuo*

A neutron diffraction study has been carried out at 2.7 ${AA}$ resolution on cubic porcine insulin at pD 6.6 using the BIX-4 at the JRR-3 reactor of JAEA. The ionization states of several amino acids in porcine insulin have been obtained at pD 6.6 and they are compared with those at pD 9 obtained by neutron diffraction as well as those at pH 6.50 and 6.98 obtained by X-ray diffraction. For the present work, a large single crystal of 2.7 mm$$^{3}$$ was obtained by dialysis. Refinement of the structure was carried out and the ${it R}$-factor was 21.4% with the free ${it R}$ being 29.4%. Protonation and deprotonation of various ionizable amino acid residues were observed and discussed on the basis of the charged states estimated by the ${it pK$_{a}$}$ values. In the case of His${it B}$5, both N$$pi$$ and N$$tau$$ of an imidazole ring are protonated at pD 6.6, but at pD 9 only N$$pi$$ is protonated. In contrast, for His${it B}$10, both N$$pi$$ and N$$tau$$ are protonated at pD 6.6 as well as at pD 9.

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パーセンタイル:43.46

分野:Chemistry, Physical

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