Thermochromatium tepidum 外周捕光天线蛋白LH2在脂质体及表面活性剂胶束中的光谱性质

doi:10.7503/cjcu20160277

高等学校化学学报 ›› 2016, Vol. 37 ›› Issue (9): 1678.doi: 10.7503/cjcu20160277

Thermochromatium tepidum 外周捕光天线蛋白LH2在脂质体及表面活性剂胶束中的光谱性质

霍一林¹, 石莹¹, 王秦玥¹, 李萝园¹, 于龙江^2,³, 王鹏¹(), 张建平¹, 王征宇²

1. 中国人民大学化学系, 北京 100872
2. 茨城大学理学部, 水户市 310-8512, 日本
3. 冈山大学自然科学学院, 冈山市700-8530, 日本

收稿日期:2016-04-25 出版日期:2016-09-10 发布日期:2016-08-26
作者简介:联系人简介: 王鹏, 男, 博士, 副教授, 主要从事天然光合体系能量转移动力学机制研究. E-mail:wpeng@iccas.ac.cn
基金资助:
国家自然科学基金(批准号: 21173265, 21273282)和中-俄合作项目(批准号: 21411130185)资助

Spectroscopic Properties of LH2 from Thermochromatium tepidum in Liposome and Detergent Micelles^†

HUO Yilin¹, SHI Ying¹, WANG Qinyue¹, LI Luoyuan¹, YU Longjiang^2,³, WANG Peng^1,^*(), ZHANG Jianping¹, WANG Zhengyu²

1. Department of Chemistry, Renmin University of China, Beijing 100872, China
2. Faculty of Science, Ibaraki University, Mito 310-8512, Japan
3. Faculty of Science, Okayama University, Okayama 700-8530, Japan

Received:2016-04-25 Online:2016-09-10 Published:2016-08-26
Contact: WANG Peng E-mail:wpeng@iccas.ac.cn
Supported by:
† Supported by the National Natural Science Foundation of China(Nos.21273282, 21173265) and the International Cooperation Project Between China and Russia(No.21411130185)

摘要/Abstract

摘要：

采用动态光散射、透射电子显微镜、紫外-可见吸收光谱和拉曼光谱对比研究了处于表面活性剂胶束和脂质体磷脂双分子层中的Thermochromatium(Tch.) tepidum LH2的光谱响应. 结果表明, 与在表面活性剂胶束中相比, 双分子层脂膜中LH2的Spirilloxanthin(一种含有13个共轭双键的类胡萝卜素)构象有明显差异; 表面活性剂及磷脂分子端基的荷电状态对B850-Qy吸收谱有显著影响; Ca²⁺结合导致B850 Qy吸收谱带红移和增色, 而H⁺结合则使该吸收谱带蓝移和减色. 对LH2脱辅基蛋白氨基酸序列的分析结果表明, Ca²⁺和H⁺的结合位点可能位于α脱辅基蛋白的C-端一侧. B850 Qy吸收谱带对Ca²⁺和H⁺的响应特性可能与Tch. tepidum适应其生存环境的能力有关.

关键词: Thermochromatium tepidum, 外周捕光天线LH2, 表面活性剂, 脂质体

Abstract:

By the use of dynamic light scattering(DLS), transmission electron microscopy(TEM), UV-Vis and resonance Raman spectroscopy, the properties of LH2 from Thermochromatium(Tch.) tepidum in two detergent micelles and in liposome were studied, respectively. The results show that in LH2-liposome the conformation of spirilloxanthin, an incorporated longer chain carotenoid, is obviously different from that in detergent micelles. The charge state of the terminal groups of detergents or lipids is the key factor affecting the B850 absorption. Generally, the calcium ions cause red-shift of the B850 absorption and hyperchromicity, while the proton has the opposite effect. Based on the amino acid sequence analysis, several amino acid residues on the C-terminus of α apo-protein were proposed to constitute the potential binding site for the calcium ion and proton. Such Ca²⁺- and H⁺-regulated changes of B850 absorption might be an adaptive mechanism to the living environment for this species.

Key words: Thermochromatium tepidum, Peripheral antenna complex LH2, Detergent, Liposome

中图分类号:

O641

TrendMD:

霍一林, 石莹, 王秦玥, 李萝园, 于龙江, 王鹏, 张建平, 王征宇. Thermochromatium tepidum 外周捕光天线蛋白LH2在脂质体及表面活性剂胶束中的光谱性质. 高等学校化学学报, 2016, 37(9): 1678.

HUO Yilin, SHI Ying, WANG Qinyue, LI Luoyuan, YU Longjiang, WANG Peng, ZHANG Jianping, WANG Zhengyu. Spectroscopic Properties of LH2 from Thermochromatium tepidum in Liposome and Detergent Micelles^†. Chem. J. Chinese Universities, 2016, 37(9): 1678.

图/表 7

Fig.1 Absorption of LH2 from Tch. tepidum in various environments at pH=8 a. in DMM; b. in LDAO; c. in liposome. The arrow shows the absorption of spirilloxanthin.

Fig.2 TEM images of LH2 from Tch. tepidum in liposome(A), DDM(B) and LDAO(C) The samples were negatively stained with 1% uranium acetate.

Fig.3 DLS data of the LH2 from Tch. tepidum in various detergents micelles or liposome systems a. in DMM(106 nm/565 nm); b. in LDAO(14 nm/257 nm); c. in liposome(73 nm). The data in the corresponding parentheses are the Rh of the vesicles for each system.

Fig.4 Resonance Raman spectra of LH2 from Tch. tepidum in various environments under the excitation at 532 nm a. in DDM; b. in LDAO; c. in liposome.

Fig.5 Qy absorption comparison of LH2 from Tch. tepidum in 0.05% DDM(a), 0.1% LDAO(b) and PC(c) liposome, under pH=8(A, C) and 3(B, D), and without Ca2+(A, B) or with 200 mmol/L Ca2+(C, D)(all the spectra are normalized at 797 nm), respectively Arrows in (A) show the splitting peaks position of B800.

Table 1 Qy absorption characteristics of LH2 from Tch. tepidum under various conditionsa

System	Without Ca²⁺				With 200 mmol/L Ca²⁺
	pH=8		pH=3		pH=8		pH=3
	λ_{max, B850}/nm	OD_B850/OD_B800	λ_{max, B850}/nm	OD_B850/OD_B800	λ_{max, B850}/nm	OD_B850/OD_B800	λ_{max, B850}/nm	OD_B850/OD_B800
In DDM	855	1.41	854	1.28	857	1.53	855	1.35
In LDAO	845	0.96	845	0.90	854	1.32	843	1.02
In liposome	850	1.05	848	1.01	854	1.33	846	1.26
In chromatophore^b	854	1.41	852	1.18	858	1.38	857	1.32

Fig.6 Molecular structure and components of detergents(A, B) or lipid(C), molecular structure and components of lipids of chromatophore from Tch. Tepidum(D) and the amino acid sequence of α-apoproteins of LH2 from Tch. Tepidum[4](E) R1—R4 are the aliphatic chain with various length and degree of unsaturation. The gray broken line indicates the B850 bacteriochlorophyll binding histidine, and the red broken line indicates the possible binding points for calcium ion or proton.

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Thermochromatium tepidum 外周捕光天线蛋白LH2在脂质体及表面活性剂胶束中的光谱性质

Spectroscopic Properties of LH2 from Thermochromatium tepidum in Liposome and Detergent Micelles^†

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