离子对人血清白蛋白影响的1H NMR研究

陈瑶; 孙鹏; 刘买利; 张许

doi:10.11938/cjmr20162527

波谱学杂志 >

2017 , Vol. 34 >Issue 3: 266 - 274

DOI: https://doi.org/10.11938/cjmr20162527

研究论文

离子对人血清白蛋白影响的¹H NMR研究

陈瑶 ,
孙鹏 ,
刘买利 ,
张许

展开

1. 波谱与原子分子物理国家重点实验室, 武汉磁共振中心(中国科学院武汉物理与数学研究所), 湖北武汉 430071;
2. 中国科学院大学, 北京 100049

收稿日期: 2016-05-10

修回日期: 2017-07-18

网络出版日期: 2017-09-05

基金资助

国家自然科学基金资助项目（21120102038，81227902，20875098，21075132）；国家重点基础研究发展计划（"973"计划）资助项目（2013CB910200）.

收起

Effects of Metal Ions on Human Serum Albumin Studied by Radiation Damping Water-Ligand Observed via Gradient Spectroscopy

CHEN Yao ,
SUN Peng ,
LIU Mai-li ,
ZHANG Xu

Expand

1. State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, National Center for Magnetic Resonance in Wuhan(Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences), Wuhan 430071, China;
2. University of Chinese Academy of Sciences, Beijing 100049, China

Received date: 2016-05-10

Revised date: 2017-07-18

Online published: 2017-09-05

Fold

摘要

人血清白蛋白（HSA）上有很多的结合位点，对药物及内源性物质的运输有很重要的作用.由于HSA分子量大且结构复杂，核磁共振（NMR）谱图信号重叠严重，因此使用常规的NMR技术很难获得天然丰度HSA上结合位点的信息.我们通过新的谱编辑技术，将横向弛豫加权（T₂W）与RD-WaterLOGSY技术相结合（即T₂W-RD-WaterLOGSY），观察到了天然丰度HSA表面的一些明显的特征信号，且这些信号的分辨率较高；通过pH滴定和2D ¹H-¹H TOCSY实验，我们对观察到的信号做了初步指认，发现部分信号来自于HSA表面的组氨酸；然后，我们通过这些特征信号研究了金属离子与HSA之间的相互作用.结果表明该技术可用于简化天然丰度HSA的谱线，在不对HSA进行突变的情况下，能反映了Zn²⁺与HSA的强结合作用以及结合位点等信息.

关键词： 核磁共振(NMR); T₂W-RD-WaterLOGSY; 相互作用; Zn²⁺; 人血清白蛋白(HSA)

本文引用格式

陈瑶 , 孙鹏 , 刘买利 , 张许 . 离子对人血清白蛋白影响的¹H NMR研究[J]. 波谱学杂志, 2017 , 34(3) : 266 -274 . DOI: 10.11938/cjmr20162527

Abstract

Human serum albumin (HSA) contains many metabolite binding sites. It has been widely studied for its functions in drug transportation. Nuclear magnetic resonance (NMR) is a frequently used tool to study HSA. However, due to its high molecular weight, the NMR spectra of HSA acquired with the conventional methods are often crowded, making spectral assignment and data interpretation difficult. In this study, a new method, which combined radiation damping water-ligand observed via gradient spectroscopy (RD-WaterLOGSY) with transverse relaxation weighted (T₂W) techniques, was proposed to simplify the NMR spectra of HSA. With the T₂W-RD-WaterLOGSY technique, the effect of pH on HSA was studied, as well as the interactions between HSA and Zn²⁺. The results showed that the relative changes in chemical shift could be used as a probe to analyze the pH changes in HSA solution and the interactions between HSA and Zn²⁺.

Key words： NMR; human serum albumin (HSA); Zn²⁺; interaction; T₂W-RD-WaterLOGSY

参考文献

[1] PETERS J R T. All about albumin:biochemistry, genetics, and medical applications[M]. America:Academic press, 1995.
[2] DUGAICZYK A, LAW S W, DENNISON O E. Nucleotide sequence and the encoded amino acids of human serum albumin mRNA[J]. P Nat Acad Sci USA, 1982, 79(1):71-75.
[3] SUGIO1 S, KASHIMA A, MOCHIZUKI S, et al. Crystal structure of human serum albumin at 2.5Å resolution[J]. Protein Eng, 1999, 12(6):439-446.
[4] HE X M, CARTER D C. Atomic structure and chemistry of human serum albumin[J]. Nature, 1992, 358(6383):209-215.
[5] STEWART P A. Modern quantitative acid-base chemistry[J]. Can J Physiol Pharm, 1983, 61(12):1444-1461.
[6] LI X, LAN W X, ZHU H, et al. Effect of pH on human blood serum studied by ¹H NMR spectroscopy[J]. Chinese J Magn Reson, 2008, 25(4):494-503. 李雪, 蓝文贤, 朱航, 等. pH对血清影响的¹H NMR研究[J]. 波谱学杂志, 2008, 25(4):494-503.
[7] COHN E J, STRONG L E, HUGHES W, et al. Preparation and properties of serum and plasma proteins. IV. A system for the separation into fractions of the protein and lipoprotein components of biological tissues and fluids1a, b, c, d[J]. J Am Chem Soc, 1946, 68(3):459-475.
[8] STEINHARDT J, KRIJN J, LEIDY J G. Differences between bovine and human serum albumins. Binding isotherms, optical rotatory dispersion, viscosity, hydrogen ion titration, and fluorescence effects[J]. Biochemistry, 1971, 10(22):4005-4015.
[9] BAL W, SOKOLOWSKA M, KUROWSKA E, et al. Binding of transition metal ions to albumin:sites, affinities and rates[J]. Biochim Biophys Acta, 2013, 1830(12):5444-5455.
[10] FERRARO G, MASSAI L, MESSORI L, et al. Cisplatin binding to human serum albumin:a structural study[J]. Chem Commun, 2015, 51(46):9436-9439.
[11] TURK B E, WONG T Y, SCHWARZENBACHER R, et al. The structural basis for substrate and inhibitor selectivity of the anthrax lethal factor[J]. Nat Struct Mol Biol, 2004, 11(11):60-66.
[12] TURK B E, WONG T Y, SCHWARZENBACHER R, et al. The structural basis for substrate and inhibitor selectivity of the anthrax lethal factor[J]. Nat Struct Mol Biol, 2004, 11(1):60-66.
[13] PETERSSON K, H KANSSON M, NILSSON H, et al. Crystal structure of a superantigen bound to MHC class Ⅱ displays zinc and peptide dependence[J]. EMBO J, 2001, 20(13):3306-3312.
[14] CAROLI S, ALIMONTI A, CONI E, et al. The assessment of reference values for elements in human biological tissues and fluids:a systematic review[J]. Crit Rev Anal Chem, 1994, 24(5,6):363-398.
[15] COUSINS R J, DUNN M A, LEINART A S, et al. Coordinate regulation of zinc metabolism and metallothionein gene expression in rats[J]. Am J Physiol Endoc M, 1986, 251(6):688-694.
[16] GIROUX E L, HENKIN R I. Macromolecular ligands of exchangeable copper, zinc and cadmium in human serum[J]. Bioinorg Chem, 1973, 2(2):125-133.
[17] PATTISON S E, COUSINS R J. Kinetics of zinc uptake and exchange by primary cultures of rat hepatocytes[J]. Am J Physiol Endoc M, 1986, 250(6):677-685.
[18] ROWE D J, BOBILYA D J. Albumin facilitates zinc acquisition by endothelial cells[J]. P Soc Exp Biol Med, 2000, 224(3):178-186.
[19] GHUMAN J, ZUNSZAIN P A, PETITPAS I, et al. Structural basis of the drug-binding specificity of human serum albumin[J]. J Mol Biol, 2005, 353(1):38-52.
[20] CARTER D C, HE X M, MUNSON S H, et al. Three-dimensional structure of human serum albumin[J]. Science, 1989, 244(4909):1195-1198.
[21] LIU T, LIU M L, JIANG L. Divalent metal ion binding to the response regulator YycFN studied by NMR spectroscopy[J]. Chinese J Magn Reson, 2016, 33(1):77-88. 刘婷, 刘买利, 姜凌. 二价金属离子与YycFN相互作用的NMR研究[J]. 波谱学杂志, 2016, 33(1):77-88.
[22] WAGNER G. An account of NMR in structural biology[J]. Nat Struct Biol, 1997, 4:841-844.
[23] CISTOLA D P, SMALL D, HAMILTON J. Carbon 13 NMR studies of saturated fatty acids bound to bovine serum albumin. I. The filling of individual fatty acid binding sites[J]. J Biol Chem, 1987, 262(23):10971-10979.
[24] CISTOLA D, SMALL D, HAMILTON J. Carbon 13 NMR studies of saturated fatty acids bound to bovine serum albumin. Ⅱ. Electrostatic interactions in individual fatty acid binding sites[J]. J Biol Chem, 1987, 262(23):10980-10985.
[25] JIANG X W, SUN P, XIAO N, et al. Sensitivity enhancement in ¹H-¹³C HSQC experiments on aromatic groups in proteins[J]. Chinese J Magn Reson, 2014, 31(1):61-70. 蒋先旺, 孙鹏, 肖楠, 等. 蛋白质芳香基团的¹H-¹³C HSQC信号增强研究[J]. 波谱学杂志, 2014, 31(1):61-70.
[26] HENZLER-WILDMAN K, KERN D. Dynamic personalities of proteins[J]. Nature, 2007, 450(7172):964-972.
[27] SNYDER D A, CHEN Y, DENISSOVA N G, et al. Comparisons of NMR spectral quality and success in crystallization demonstrate that NMR and X-ray crystallography are complementary methods for small protein structure determination[J]. J Am Chem Soc, 2005, 127(47):16505-16511.
[28] LIN D H, HONG J. Mapping protein-ligand interaction by NMR techniques[J]. Chinese J Magn Reson. 2005, 22(3):321-341. 林东海, 洪晶. 用NMR技术研究蛋白质-配体相互作用[J]. 波谱学杂志, 2005, 22(3):321-341.
[29] BLINDAUER C A, HARVEY I, BUNYAN K E, et al. Structure, properties, and engineering of the major zinc binding site on human albumin[J]. J Biol Chem, 2009, 284(34):23116-23124.
[30] DALVIT C, FOGLIATTO G, STEWART A, et al. WaterLOGSY as a method for primary NMR screening:practical aspects and range of applicability[J]. J Biomol NMR, 2001, 21(4):349-359.
[31] SUN P, JIANG X W, JIANG B, et al. Biomolecular ligands screening using radiation damping difference WaterLOGSY spectroscopy[J]. J Biomol NMR, 2013, 56(3):285-290.
[32] GRZESIEK S, BAX A. Measurement of amide proton exchange rates and NOEs with water in 13C/15N-enriched calcineurin B[J]. J Biomol NMR, 1993, 3(6):627-638.
[33] LIU M L, MAO X A, YE C H, et al. Improved WATERGATE pulse sequences for solvent suppression in NMR spectroscopy[J]. J Magn Reson, 1998, 132(1):125-129.
[34] WANG J, ZHANG X, SUN P, et al. The impact of pulse duration on composite WATERGATE pulse[J]. J Magn Reson, 2010, 206(2):205-209.
[35] STEWART A J, BLINDAUER C A, BEREZENKO S, et al. Interdomain zinc site on human albumin[J]. P Nat Acad Sci USA, 2003, 100(7):3701-3706.

Options

文章导航

模态框（Modal）标题

摘要

本文引用格式

Abstract

参考文献