Functionality Research of Keggin-type Polyoxotungstate as Potent Mushroom Tyrosinase Inhibitors†

doi:10.7503/cjcu20131010

Abstract

Abstract:

H₃PW₁₂O₄₀ and H₄SiW₁₂O₄₀(abbreviated as PW₁₂ and SiW₁₂) were taken as inhibitors and their inhibitory effects and mechanisms on mushroom tyrosinase were evaluated by Native polyacrylamide gel electrophoresis(PAGE) electrophoresis method and kinetic enzymatic method. The Native-PAGE electrophoresis results show that tyrosinase are encoded by a multigene family, their molecular weight are mainly 3×10⁴—3.4×10⁴, 4.2×10⁴—4.6×10⁴ and 6.4×10⁴—6.8×10⁴, which were all active.The inhibitory effects of Kiggin-type polyoxotungstates, H₃PW₁₂O₄₀ and H₄SiW₁₂O₄₀ on mushroom tyrosinase were evaluated, and then the inhibitory mechanisms were further studied combined with the kinetic enzymatic method. The results indicated that the enzyme was inactive when the concentration of PW₁₂ and SiW₁₂ was 13 and 25 mmol/L, respectively. More-over, the kinetic enzymatic method suggested that both PW₁₂ and SiW₁₂ exhibited potent inhibitory activities with IC₅₀ value of 1.57 and 2.31 mmol/L, respectively. PW₁₂ was found to be a mixed-type inhibitor with K_I=0.34 mmol/L and K_IS=0.43 mmol/L, while SiW₁₂ was suggested to be competitive inhibitor of tyrosinase with K_I of 0.59 mmol/L. Based on a comprehensive consideration of IC₅₀ and inhibitory constants, PW₁₂ was more effective against mushroom tyrosinase than SiW₁₂.

Key words: Polyoxometalate, Mushroom tyrosinase, Native polyacrylamide gel electrophoresis(PAGE), Inhibition effect

CLC Number:

O614.5
Q55

TrendMD:

ZHENG Aping, WANG Fang, XING Rui, JIANG Aihua, WANG Li. Functionality Research of Keggin-type Polyoxotungstate as Potent Mushroom Tyrosinase Inhibitors^†[J]. Chem. J. Chinese Universities, 2014, 35(3): 476.

Figures/Tables 6

Fig.1 Electrophoresis analysis of protein mole-cular weight and species of mushroom tyrosinaseLane M: protein molecular weight markers; lane 1: tyrosinase stained with coomassie blue; lane 2, 3: tyrosinase stained with L-DOPA.

Fig.2 Native-PAGE analysis of Inhibitory effects of PW12(A) and SiW12(B) on the mushroom tyrosinaseConcentrations of PW12 for lanes 1—10 were 0, 5, 10, 11, 12, 13, 14, 15, 16, 18 mmol/L, respectively. Concentrations of SiW12 for lanes 1—10 were 0, 5, 10, 15, 16, 17, 18, 19, 20, 25 mmol/L, respectively.

Fig.3 Inhibitory effects of PW12(a) and SiW12(b) on the mushroom tyrosinase

Fig.4 Inhibitory mechanism of PW12(A) and SiW12(B) on the mushroom tyrosinase (A) Concentrations of curves a—c were 0, 0.67 and 1 mmol/L, respectively; (B) concentrations of curves a—c were 0, 1.67 and 3.33 mmol/L, respectively.

Fig.5 Inhibitory types and the inhibitory constants of PW12 on mushroom tyrosinase for the catalysis of L-DOPA(A), the secondary plot of the slope vs. the concentrations of PW12 for determining the inhibitory constant KI(B), the secondary plot of the intercept vs. the concentrations of PW12 for determining the inhibitory constant KIS(C)(A) Concentrations of PW12 for curves a—d were 0, 0.67, 1 and 1.33 mmol/L, respectively.

Fig.6 Inhibitory types and the inhibitory constants of SiW12 on mushroom tyrosinase for the catalysis of L-DOPA(A) and the secondary plot of the intercept vs. the concentrations of PW12 for determining the inhibitory constant KIS(B)(A) Concentrations of SiW12 for curves a—d were 0, 0.67, 1.33 and 2 mmol/L, respectively.

References 42

[1]	Parenicová L., Enzymes in Fruit and Vegetable Processing, CRC Press, New York, 2010, 20—22
[2]	De Castro E., Barrett D. M., Jobling J., Mitcham E. J., Postharvest Biol. Tec., 2008, 48(2), 182—191
[3]	Bao K., Dai Y., Zhu Z. B., Tu F. J., Zhang W. G., Yao X. S., Bioorg. Med. Chem., 2010, 18(18), 6708—6714
[4]	Lin L., Dong Y., Zhao H., Wen L., Yang B., Zhao M., Food Chem., 2011, 129(3), 884—889
[5]	Mayer A. M., Phytochemistry, 2006, 67(21), 2318—2331
[6]	Li Z. C., Chen L. H., Yu X. J., Hu Y. H., Song K. K., Zhou X. W., Chen Q. X., J. Agric. Food Chem., 2010, 58(23), 12537—12540
[7]	Ha Y. M., Kim J. A., Park Y. J., Park D., Kim J. M., Chung K. W., Lee E. K., Park J. Y., Lee J. Y., Lee H. J., Yoon J. H., Moon H. R., Chung H. Y., BBA-General Subjects,2011, 1810(6), 612—619
[8]	Morrison M. E., Yagi M. J., Cohen G., Proc. Natl. Acad. Sci., 1985, 82(9), 2960—2964
[9]	Takahashi S., Kamiya T., Saeki K., Nezu T., Takeuchi S. I., Takasawa R., Sunaga S., Yoshimori A., Ebizuka S., Abe T., Tanuma S. I., Bioorg. Med. Chem., 2010, 18(22), 8112—8118
[10]	Zhang J. P., Chen Q. X., Song K. K., Xie J. J., Food Chem., 2006, 95(4), 579—584
[11]	Yi W., Cao R., Wen H., Yan Q., Zhou B. H., Wan Y. H., Ma L., Song H., Bioorg. Med. Chem. Lett., 2008, 18(24), 6490—6493
[12]	Lou S. N., Yu M. W., Ho C. T., Food Chem., 2012, 135(3), 1091—1096
[13]	Liu J., Wu F., Chen L., Zhao L., Zhao Z., Wang M., Lei S., Food Chem., 2012, 135(4), 2872—2878
[14]	Song Y. M., Ha Y. M., Kim J. A., Chung K. W., Uehara Y., Lee K. J., Chun P., Byon Y., Chung H. Y., Moon H. R., Bioorg. Med. Chem. Lett., 2012, 22(24), 7451—7455
[15]	Sun Z., Li F., Xu L., Liu S., Zhao M., Xu B., J. Phys. Chem. C,2012, 116(10), 6420—6426
[16]	Huang P., Qin C., Su Z. M., Xing Y., Wang X. L., Shao K. Z., Lan Y. Q., Wang E. B., J. Am. Chem. Soc., 2012, 134(34), 14004—14010
[17]	Chen B. N., Feng Z. G., Wang L., Chem. J. Chinese Universities,2009, 30(11), 2136—2138
	(陈丙年, 冯珍鸽, 王力. 高等学校化学学报, 2011, 32(5), 1033—1036)
[18]	Wall M. J., Wigmore G., Lopatár J., Frenguelli B. G., Dale N., Neuropharmacology, 2008, 55(7), 1251—1258
[19]	Sun X., Wu Y., Gao W., Enjyoji K., Csizmadia E., Müller C. E., Murakami T., Robson S. C., Gastroenterology,2010, 139(3), 1030—1040
[20]	Prudent R., Moucadel V., Laudet B., Barette C., Lafanechère L., Hasenknopf B., Li J., Bareyt S., Lacote E., Thorimbert S., Malacria M., Gouzerh P., Cochet C., J. Chembiol., 2008, 15(7), 683—692
[21]	Fraqueza G., Ohlin C. A., Casey W. H., Aureliano M., J. Inorgan. Biochem., 2012, 107(1), 82—89
[22]	Turner T. L., Nguyen V. H., McLauchlan C. C., Dymon Z., Dorsey B. M., Hooker J. D., Jones M. A.,J. Inorgan. Biochem., 2012, 108(3), 96—104
[23]	Flütsch A., Schroeder T., Grütter M. G., Patzke G. R., Bioorg. Med. Chem. Lett., 2011, 21(4), 1162—1166
[24]	Geng J., Li M., Ren J., Wang E., Qu X., Angew. Chem. Int. Ed., 2011, 123(18), 4270—4274
[25]	Wu Q., Wang J., Zhang L., Hong A., Ren J., Angew. Chem. Int. Ed., 2005, 117(26), 4116—4120
[26]	Stephan H., Kubeil M., Emmerling F., Müller C. E., Eur. J. Inorg. Chem., 2013, 2013(10/11), 1585—1594
[27]	Schägger H., Nat. Prot., 2006, 1(1), 16—22
[28]	Xie L. P., Chen Q. X., Huang H., Liu X. D., Chen H. T., Zhang R. Q., Int. J. Biochem. Cell. B,2003, 35(12), 1658—1666
[29]	Yi W., Cao R., Peng W., Wen H., Yan Q., Zhou B., Ma L., Song H., Eur. J. Med. Chem., 2010, 45(2), 639—646
[30]	Zhu Y. J., Zhou H. T., Hu Y. H., Tang J. Y., Su M. X., Guo Y. J., Chen Q. X., Liu B., Food Chem., 2011, 124(1), 298—302
[31]	Richter H., Lieberei R., Strnad M., Novák O., Gruz J., Rensing S. A., von Schwartzenberg K., J. Exp. Bot., 2012, 63(14), 5121—5135
[32]	Yamamoto H., Kudo T., Masuko N., Miura H., Sato S., Tanaka M., Tanaka S., Takeuchi S., Shibahara S., Takeuchi T., Pigm. Cell. Res., 1992, 5(5), 284—294
[33]	Bernan V., Filpula D., Herber W., Bibb M., Katz E., Gene,1985, 37(1), 101—110
[34]	Gerdemann C., Eicken C., Krebs B., Accounts of Chemical Research,2002, 35(3), 183—191
[35]	Wichers H. J., Recourt K., Hendriks M., Ebbelaar C. E. M., Biancone G., Hoeberichts F. A., Mooibroek H., Soler-Rivas C., Appl. Microbiol. Biot., 2003, 61(4), 336—341
[36]	Wu J., Chen H., Gao J., Liu X., Cheng W., Ma X., Biotech. Lett., 2010, 32(10), 1439—1447
[37]	Han P., Chen C. Q., Zhang C. L., Song K. K., Zhou H. T., Chen Q. X., Food Chem., 2008, 107(2), 797—803
[38]	Yi W., Wu X., Cao R., Song H., Ma L., Food Chem., 2009, 117(3), 381—386
[39]	Huang X. H., Chen Q. X., Wang Q., Song K. K., Wang J., Sha L., Guan X., Food Chem., 2006, 94(1), 1—6
[40]	Yi W., Cao R., Wen H., Yan Q., Zhou B., Ma L., Song H.,Bioorg. Med. Chem. Lett., 2009, (19), 6157—6160
[41]	Wang E.B., Hu C. W., Xu L., Introduction of Polyoxometalate Chemistry, Chemical Industry Press, Beijing, 1998, 171—184
	(王恩波, 胡长文, 许林. 多酸化学导论. 北京: 化学工业出版社, 1998, 171—184)
[42]	Yamase T., Mol. Eng., 1993, 3(1—3), 241—262