Kinetic Study of the Interaction Between Lactose-containing Assembly and Protein Through Dynamic Light Scattering Technique†

doi:10.7503/cjcu20180367

Abstract

Abstract:

Non-covalently connected micelles(NCCM) strategy was employed to obtain glyco-micelles covered by sugars in a convenient way. The impact factors for the recognition between carbohydrate and protein in solution were studied using dynamic light scattering(DLS) in real time. The results indicate that the agglutination ability of different lectins is of significant distinction, as measured by the formation rate of aggregates. For glyco-micelles covered by 100% lactose, the binding rates to different lectins can be recorded as SBA?LecA≈Galectin 3> ECA≈Galectin 1, which suggests that not only number of binding sites but also how they are displayed have great influence on the kinetics of the multivalent interaction. Meanwhile, Human Galectin 3, a protein of important biological function with rather mysterious binding behavior, was studied further as an application of the method.

Key words: Glycopolymer, Lectin, Dynamic light scattering, Multivalent interaction

CLC Number:

O631

TrendMD:

ZHANG Pan,SUN Pengfei,LIN Mingchang,GAO Zhenfei,CHEN Guosong. Kinetic Study of the Interaction Between Lactose-containing Assembly and Protein Through Dynamic Light Scattering Technique^†[J]. Chem. J. Chinese Universities, 2019, 40(1): 160.

Figures/Tables 9

Table 1 Mass fraction(%) of the shell and core of glyco-micelles

Sample	PGlc(shell)	PLac(shell)	PtBMA(core)
Lac-0	100	0	100
Lac-25	75	25	100
Lac-50	50	50	100
Lac-75	25	75	100
Lac-100	0	100	100

Fig.1 Structures of PtBMA-co-PBOB and glycopolymers PLac and PGlc

Fig.2 TEM image of Lac-100(A) and <Rh> of Lac-100(B)

Table 2 Hydrodynamic radius of glyco-micelles with different coverage degree of lactose*

Sample	<R_h>/nm	PDI
Lac-0	63	0.08
Lac-25	60	0.22
Lac-50	72	0.28
Lac-75	80	0.38
Lac-100	60	0.10

Fig.3 <Rh> of Lac-100 and Lac-0 with titration of SBA(A) and <Rh> distribution of Lac-100 with titration of SBA(B)Sidenotes are the volume of SBA added to the solution. Mean value of three continuous experiments. Concentration of glyco-micelles is 0.04 mg/mL.

Fig.4 <Rh> of Lac-100 with the titration of different lectinsc(Lac-100): 0.04 mg/mL.

Fig.5 Crystal structure of SBA complexed with 2,6-pentasaccharide(PDB code 2SBA)(A) and pseudomonas aeruginosa lectin(LecA) complexed with GalA-QRS(PDB code 4LKD)(B)

Fig.6 <Rh> of glyco-micelles with titration of different lectins(A) Lac-25; (B) Lac-50; (C) Lac-75; (D) Lac-100.

Fig.7 <Rh> of glyco-micelles with titration of Galectin 3

References 25

[1]	Dwek R. A., Chem. Rev., 1995, 23(1), 1—25
[2]	Imberty A., Varrot A., Curr. Opin. Struct. Biol.2008, 18(5), 567—576
[3]	Huang Y., Huang J.H., Xie Q. J., Yao S. Z.,Prog. Chem., 2008, (6), 942—950
	(黄毅, 黄金花, 谢青季, 姚守拙. 化学进展, 2008, (6), 942—950)
[4]	Pang P. C., Dell A., Science,2011, 333(6050), 1761—1764
[5]	Lis H., Sharon N., Chem. Rev.,1998, 98(2), 637—674
[6]	Godula K., Bertozzi C. R., J. Am. Chem. Soc., 2012, 134(38), 15732—42
[7]	Liang P. H., Wang S. K., Wong C. H., J. Am. Chem. Soc., 2007, 129(36), 11177—84
[8]	He X. P., Wang X. W., Jin X. P., J. Am. Chem. Soc., 2011, 133(10), 3649—3657
[9]	Wan A. J., Wang K., Zhang H. C., Li H. L., Wang D. N., Chinese J. Anal. Chem., 2012, 40(11), 1780—1788
	(万锕俊, 王琨, 张洪才, 李慧丽, 王德农. 分析化学, 2012, 40(11), 1780—1788)
[10]	Zhao X.L., The Interactions of Marine Polysaccharides and Their Derivatives with Proteins by Carbohydrate Microarray, Ocean University of China, Qingdao, 2013
	(赵小亮. 利用糖芯片研究海洋多糖及其衍生物与蛋白质的相互作用,青岛: 中国海洋大学, 2013)
[11]	Su L., Zhao Y., Chen G. S., Polym. Chem., 2012, 3(6), 1560—1566
[12]	Sun P. F., Lin M. C., Chen G. S., Sci. Chinese Chem.,2016, 59(12), 1—5
[13]	Lin M.C., Chen G. S., Acta Polym. Sin., 2017, (7), 1113—1120
	(林铭昌, 陈国颂. 高分子学报, 2017, (7), 1113—1120)
[14]	Lin M. C., Sun P. F., Chen G. S., Chem. Commun., 2014, 50(68), 9779—9782
[15]	Zhu H., Yuan X.F., Zhao H. Y., Liu S. Y., Jiang M.,Chinese J. Appl. Chem., 2001, (5), 336—341
	(朱蕙, 袁晓凤, 赵汉英, 刘世勇, 江明. 应用化学, 2001, (5), 336—341)
[16]	Guo M. Y., Jiang M., Soft Matter.,2009, 5(3), 495—500
[17]	Christopher W. C., Jason E. G., Motomu K., J. Am. Chem. Soc., 2002, 124(8), 1615—1619
[18]	Nieminen J., Kuno A., Hirabayashi J., J. Biol. Chem.,2007, 282(2), 1374—1383
[19]	Adriana L., Emma S., Nilsson U. J., J. Biol. Chem., 2012, 287(26), 21751—56
[20]	De B. H., Lis H., Van T. H., J. Biol. Chem., 1984, 259(11), 7067—7074
[21]	Kadam R. U., Myriam B., Matthew H., Angew. Chem. Int. Ed.,2011, 50(45), 10631—35
[22]	Iglesias J. L., Lis H., Sharon N., Eur. J. Biochem.,1982, 123(2), 247—252
[23]	Camby I., Mercier M. L., Lefranc F., Glycobiology,2006, 16(11), 137—138
[24]	Li M. M., Xiong Y. T., Qing G. Y., Sun T. L., Prog. Biochem. Biophys., 2016, 43(2), 115—127
	(李闵闵, 熊雨婷, 卿光焱, 孙涛垒. 生物化学与生物物理进展, 2016, 43(2), 115—127)
[25]	Di L. S., Sundblad V., Cerliani J. P., Biochemistry,2011, 50(37), 7842—7857