Mass Spectrometric Analysis of N-Glycans of Glycoprotein Separated by SDS-PAGE Gel from Ginkgo Seed†

doi:10.7503/cjcu20180456

Abstract

Abstract:

The glycoproteins in Ginkgo seeds were separated and isolated by sodium dodecyl sulfate polyacry-lamide gel electrophoresis(SDS-PAGE). The N-glycans released from glycoproteins were qualitatively and quantitatively analyzed by electrospray ionization mass spectrometry(ESI-MS) and on-line liquid chromatography-mass spectrometry(LC-UV-MS/MS²). As a result, eleven glycoproteins containing oligomannose type glycans(4.88%) and complex type glycans(95.12%) were obtained. And most of the N-glycans were modified by the core α-1, 3- and β-1, 2-xylose in the 21000, 36000 and 50000 glycoproteins, which accounted for 68.23%, 64.37% and 75.09%, respectively. This study can highlight the understanding of protein glycosylation in Ginkgo and provide foundation in the research of Ginkgo biloba seed allergens.

Key words: Ginkgo glycoprotein, Sodium dodecyl sulfate polyacrylamide gel electrophoresis, PNGase F, Aqueous ammonia, Electrospray ionization mass spectrometry

CLC Number:

TrendMD:

LI Cheng,WANG Chengjian,JIN Wanjun,HAN Jianli,YANG Meifang,GAO Xi,HUANG Linjuan,WANG Zhongfu. Mass Spectrometric Analysis of N-Glycans of Glycoprotein Separated by SDS-PAGE Gel from Ginkgo Seed^†[J]. Chem. J. Chinese Universities, 2019, 40(1): 69.

Figures/Tables 6

Fig.1 SDS-PAGE profile dyed by G250(A) and PAS(B) of Ginkgo total protein

Fig.2 ESI-MS spectra of the N-glycan released from Ginkgo total protein by PNGase F

Fig.3 ESI-MS spectra of the N-glycan released from Ginkgo total protein by aqueous ammonia(A) and its glycan labeled with PMP(B)

Fig.4 Extracted ion chromatograms(EICs) of the N-glycans labeled with PMP of Ginkgo total protein

Fig.5 Quantitative distribution of the N-glycans from Ginkgo total proteinXM: xylose-modified type; XFM: xylose and fucose-modified type.

Fig.6 ESI-MS spectra of the N-glycans of protein bands

References 27

[1]	Ruhaak L. R., Kim K., Stroble C., Taylor S. L., Hong Q., Suzanne M. S., Carlito B., Lebrilla C. B., Leiserowitz G., J. Proteome Res., 2016, 15(3), 1002—1010
[2]	Hart G. W., Copeland R. J., Cell,2010, 143(5), 672—676
[3]	Pang P. C., Chiu P. C., Lee C. L., Chang L. Y., Panico M., Dell A., Science,2011, 333(6050), 1761—1764
[4]	Gu J., Ji T. I., Xu Q., Kariya Y., Gu W., Fukuda T., Du Y., Glycoconjugate J., 2012, 29(8), 599—607
[5]	Altmann F., Int. Arch. Allergy Imm., 2007, 142(2), 99—115
[6]	Li M., Jin Y., Wang X., Wu Q., Liu Y., Li P., Li X., Chem. Res. Chinese Universities,2016, 32(6), 943—946
[7]	Bublin M., Radauer C., Wilson I. B., Kraft D., Scheiner O., Breiteneder H., Hoffmann-Sommergruber K., FASEB J., 2003, 17(12), 1697—1699
[8]	Wilson I. B., Curr. Opin. Struc. Biol., 2002, 12(5), 569—577
[9]	Du C., Huang Y., J. Dermatol. Venereol.,2015, 37(5), 301—302
	(杜策, 黄莹. 皮肤病与性病, 2015, 37(5), 301—302)
[10]	Yang J. T., Wu C. E., Li Y. Y., Jia S. Q., Fan G. J., Pan H. M., Agr. Sci. China,2010, 43(17), 3616—3623
	(杨剑婷, 吴彩娥, 李莹莹, 贾韶千, 范龚健, 潘红梅. 中国农业科学, 2010, 43(17), 3616—3623)
[11]	Fötisch K., Vieths S., Glycoconjugate J., 2001, 18(5), 373—390
[12]	Mari A., Int. Arch. Allergy Imm., 2002, 129(4), 286—295
[13]	Foetisch K., Westphal S., Lauer I., Retzek M., Altmann F. A., Kolarich D., Scheurer S., Vieths S., J. Allergy Clin. Immuun., 2003, 111(4), 889—896
[14]	Shi X., Zhang T., Li X., Feng Y., Tan X., Jin Y., Chem. Res. Chinese Universities,2016, 32(4), 556—560
[15]	Paulus K. E., Mahler V., Pabst M., Kogel K. H., Altmann F., Sonnewald U., Front. Plant Sci., 2011, 2, 42
[16]	Okano M., Kimura Y., Kino K., Michigami Y., Sakamoto S., Sugata Y., Maeda M., Matsuda F., Kimura M., Ogawa T., Nishizaki K., Clin. Exp. Allergy,2004, 34(5), 770—778
[17]	Maeda M., Takeda N., Mano A., Yamanishi M., Kimura M., Kimura Y., Biosci. Biotech. Bioch., 2013, 77(6), 1269—1274
[18]	Li Y. Y., Wu C. E., Yang J. T., Jia S. Q., Xu W. B., Peng F. R., Food Sci., 2010, 31(22), 36—40
	(李莹莹, 吴彩娥, 杨剑婷, 贾韶千, 徐文斌, 彭方仁. 食品科学, 2010, 31(22), 36—40)
[19]	Neville Jr D. M., Glossmann H., Method Enzymol., 1974, 32, 92—102
[20]	Wang C. J., Lu Y., Han J. L., Jin W. J., Li L. M., Zhang Y., Song X. Z., Huang L. J., Wang Z. F., J. Prothome Res., 2018, 17(7), 2345—2357
[21]	Honda S., Akao E., Suzuki S., Okuda M., Kakehi K., Nakamura J., Anal. Biochem.,1989, 180(2), 351—357
[22]	James A. T., Yang A., Food Chem., 2016, 194, 284—289
[23]	Liu C., Wang H., Cui Z., He X., Wang X., Zeng X., Ma H., Food Chem., 2007, 102(4), 1310—1316
[24]	Li Y. Y., Wu C. E., Yang J. T., Fan G. J., Food Sci., 2011, 32(7), 16—20
	(李莹莹, 吴彩娥, 杨剑婷, 范龚健. 食品科学, 2011, 32(7), 16—20)
[25]	Aalberse R. C., Koshte V., Clemens J. G., J. Allergy Clin. Immuun., 1981, 68(5), 356—364
[26]	Tretter V., Altmann F., Kubelka V., Int. Arch. Allergy Imm., 1993, 102(3), 259—266
[27]	Ree R. V., Cabanesmacheteau M., Akkerdaas J., J. Biol. Chem., 2000, 275(15), 11451—11458