金属离子对豆腐果苷与牛血清白蛋白相互作用的影响

doi:10.3969/j.issn.0253-2417.2019.01.017

林产化学与工业 ›› 2019, Vol. 39 ›› Issue (1): 115-122.doi: 10.3969/j.issn.0253-2417.2019.01.017

金属离子对豆腐果苷与牛血清白蛋白相互作用的影响

赵丹丹¹,金小伟¹,石婧楠¹,张玉贞¹,边贺东^1,^2,*(),廖永志²

1. 广西民族大学化学化工学院；广西林产化学与工程重点实验室，广西南宁 530006
2. 广西水产科学研究院，广西南宁 530021

收稿日期:2018-09-30 出版日期:2019-02-25 发布日期:2019-03-14
通讯作者: 边贺东 E-mail:gxunchem@163.com
作者简介:赵丹丹(1993—)，女，山东济南人，硕士生，研究方向为生物无机化学
基金资助:
广西自然科学基金资助项目(2016GXNSFDA380005);广西科技基地和人才专项(2017AD19029);广西民族大学研究生教育创新计划资助项目(chxps201820)

Effect of Metal Ions on Interaction Between Helicid and Bovine Serum Albumin

Dandan ZHAO¹,Xiaowei JIN¹,Jingnan SHI¹,Yuzhen ZHANG¹,Hedong BIAN^1,^2,*(),Yongzhi LIAO²

1. School of Chemistry and Chemical Engineering, Guangxi University for Nationalities; Guangxi Key Laboratory of Chemistry and Engineering of Forest Products, Nanning 530006, China
2. Guangxi Academy of Fishery Sciences, Nanning 530021, China

Received:2018-09-30 Online:2019-02-25 Published:2019-03-14
Contact: Hedong BIAN E-mail:gxunchem@163.com
Supported by:
广西自然科学基金资助项目(2016GXNSFDA380005);广西科技基地和人才专项(2017AD19029);广西民族大学研究生教育创新计划资助项目(chxps201820)

摘要/Abstract

摘要：

在模拟人体生理环境的条件下，运用荧光光谱法研究了豆腐果苷(HLD)与牛血清白蛋白(BSA)的相互作用，以及不同金属离子的加入对HLD-BSA体系产生的影响。研究结果表明：HLD与BSA发生了静态猝灭，形成了稳定的化合物。通过Stern-Volmer方程计算可知，HLD与BSA在293、298和303 K的猝灭常数分别为6.316、5.147和4.040×10¹² L/(mol·s)，结合常数分别为9.10×10⁵、7.76×10⁵和4.97×10⁵ L/mol。非辐射能量转移Föster理论分析表明：BSA中色氨酸残基与HLD的作用距离(r)为4.79 nm，当加入金属离子后，r值均有不同程度的降低；热力学分析表明：HLD和BSA之间的作用力以氢键和范德华力为主。运用圆二色光谱研究了HLD和金属离子的加入对BSA二级结构的影响，结果表明：HLD的加入导致了BSA的α-螺旋结构百分比的下降，金属离子的存在进一步诱导了BSA二级结构的变化，BSA的α-螺旋结构百分比明显下降。

关键词: 牛血清白蛋白, 豆腐果苷, 金属离子, 相互作用

Abstract:

The effect of Fe³⁺, Cu², Ni²⁺, Zn²⁺on the interaction reaction between helicid(HLD)and bovine serum albumin(BSA)was investigated by fluorescence spectroscopy under similarity physiological conditions. The results showed that the fluorescence of BSA was quenched via HLD by forming stable complexes. The quenching constants(k_q)calculated by Stern-Volmer equation under 293, 298 and 303 K were 6.316, 5.147 and 4.040×10¹² L/(mol·s), respectively. The binding constants(K) were 9.10×10⁵, 7.76×10⁵, and 4.97 ×10⁵ L/mol, respectively. The distance between the tryptophan residues of BSA and HLD was 4.79 nm by using Föster′s equation, and it decreased after interaction with metal ions. The theromodynamic parameters indicated that the force between HLD and BSA were hydrogen band and van der Waals forces. The effects of HLD and different metal ions on the secondary structure of BSA were studied using circular dichroism(CD) spectroscopy. The results showed that the addition of HLD changed the conformation of BSA in the binding reaction, and α-helix content decreased. The secondary structure of BSA was also changed by the addition of metal ions and α-helix content further reduced obviously.

Key words: bovine serum albumin, helicid, metal ions, interaction

中图分类号:

TQ35

赵丹丹,金小伟,石婧楠,张玉贞,边贺东,廖永志. 金属离子对豆腐果苷与牛血清白蛋白相互作用的影响[J]. 林产化学与工业, 2019, 39(1): 115-122.

Dandan ZHAO,Xiaowei JIN,Jingnan SHI,Yuzhen ZHANG,Hedong BIAN,Yongzhi LIAO. Effect of Metal Ions on Interaction Between Helicid and Bovine Serum Albumin[J]. Chemistry and Industry of Forest Products, 2019, 39(1): 115-122.

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T/K	体系systems	10^-4K_SV/(L·mol^-1)	10^-12kq/(L·mol^-1·s^-1)	R
293	HLD-BSA	6.316	6.316	0.9867
	HLD-BSA	5.147	5.147	0.9866
	HLD-BSA-Fe³⁺	7.874	7.874	0.9935
298	HLD-BSA-Cu²⁺	6.116	6.116	0.9957
	HLD-BSA-Zn²⁺	5.131	5.131	0.9975
	HLD-BSA-Ni²⁺	9.385	9.385	0.9832
303	HLD-BSA	4.040	4.040	0.9834

T/K	体系systems	10^-5K/(L·mol^-1)	f_a	R
293	HLD-BSA	9.10	0.36	0.9955
	HLD-BSA	7.76	0.35	0.9908
	HLD-BSA-Fe³⁺	2.68	0.56	0.9912
298	HLD-BSA-Cu²⁺	2.16	0.54	0.9970
	HLD-BSA-Zn²⁺	2.32	0.56	0.9990
	HLD-BSA-Ni²⁺	2.84	0.61	0.9922
303	HLD-BSA	4.97	0.34	0.9842

体系system	J/(×10^-14 cm³·L·mol^-1)	R₀/nm	E	r/nm
BSA-HLD-Fe³⁺	6.34	3.35	0.186	4.48
BSA-HLD-Cu²⁺	6.15	3.35	0.097	4.37
BSA-HLD-Zn²⁺	6.83	3.35	0.088	4.58
BSA-HLD-Ni²⁺	6.64	3.35	0.079	4.66

T/K	K/(×10⁵ L·mol^-1)	ΔG^{${\rm{\rlap{-} o}}$}/(kJ·mol^-1)	ΔH^{${\rm{\rlap{-} o}}$}/(kJ·mol^-1)	ΔS^{${\rm{\rlap{-} o}}$}/(J·mol^-1K^-1)
293	9.14	-134.71
298	7.49	-119.42	-224.281	-673.17
303	4.97	-101.02

C_M:C_HLD:C_BSA	Fe³⁺-BSA-HLD/%	Cu²⁺-BSA-HLD/%	Zn²⁺-BSA-HLD/%	Ni²⁺-BSA-HLD/%
0:0:1	40.25	40.25	40.25	40.25
0:1:1	40.03	40.03	40.03	40.03
1:1:1	39.30	39.70	38.39	38.60
2:1:1	37.93	37.66	37.64	38.01

金属离子对豆腐果苷与牛血清白蛋白相互作用的影响

Effect of Metal Ions on Interaction Between Helicid and Bovine Serum Albumin

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