欢迎来到糖化学与生物技术教育部重点实验室
细胞工程课题组
  2017-05-03  

 

长简介
姓名:           高晓冬 (Xiao-Dong Gao)

出生年月:    1965年1月14日
职称:           教授,博士生导师

研究领域:    细胞分子生物学、糖生物学、生物工程
人才称号:    江苏省双创人才
E-mail:        xdgao@jiangnan.edu.cn/

教育背景
1990-1994     日本 东京大学农学研究科 获博士学位
1988-1990     日本 山梨大学工学研究科 获硕士学位
1984-1988     日本 山梨大学工学部 获学士学位

工作经历: 
2011-至今      江南大学生物工程学院 教授
2006-2011     日本 北海道大学 先端生命科学研究院 副教授 博士生导师
2003-2006     日本 国立产业综合研究所(AIST)糖锁研究中心主任研究员
2002-2003     美国 纽约州立大学 助理教授
1998-2002     美国 纽约州立大学 博士后研究员
1995-1998     日本 国立生命科学和技术研究所(NIBH)博士后

                                                                                                                                                             

研究队伍
姓名:          王宁(Ning Wang)   
职称:           副研究员
研究領域:    糖生物学、糖化学生物学
E-mail:        wangning@jiangnan.edu.cn    

姓名:         李子杰(Zijie Li)      
职称:         副教授
E-mail:       lizijie@jiangnan.edu.cn


主要研究背景及目标

背景介绍

   N-糖基化是真核细胞蛋白质翻译后修饰的主要形式之一,它开始于内质网膜上的多萜醇寡糖前体DLO(dolichol-linked oligosaccharide)的合成。DLO由14个单糖组成,它们的修饰分别由13个糖基转移酶(Alg蛋白)来承担。DLO合成首先在内质网膜的胞质侧举行,随即被多萜醇寡糖翻转酶(flipase)把中间产物M5-Dol(Man5GlcNAc2-PP-Dol)从内质网膜外翻转到腔内,由腔内Alg糖基转移酶继续完成余下的修饰反应,生成最终产物Glc3Man9GlcNAc2-PP-Dol(图1)。最终产物的核心寡糖部分将被寡糖转移酶复合体OST(oligosaccharyltransferase complex)从多萜醇二磷酸上切开,转移到新翻译生成肽链的Asn-X-Ser/Thr序列中的天冬酰胺残基上形成N-糖苷键,从此开始它作为蛋白质N-聚糖的生涯。

   前期的研究积累证明内质网N-糖基化途径在各类真核细胞中保守性较高,酵母、植物和动物细胞都共有一致的Alg糖基转移酶和OST寡糖转移酶甚至M5-Dol翻转酶体系。但是,在一些寄生虫细胞(如:T. gondii; E. histolytica)中也发现了部分Alg糖基转移酶、OST寡糖转移酶和M5-Dol翻转酶体系相关基因的缺损现象,说明了内质网N-糖基化途径的多样性和复杂性。这些特征奠定了真核细胞N-糖基化修饰的研究以及其工业应用技术开发的基础。近期的研究结果显示DLO合成途径前期(胞质侧)的Alg糖基转移酶具有调控整个细胞N-糖基化的功能,但具体分子机制还有待阐明。另外,M5-Dol翻转酶至今尚未被发现,翻转机制仍处于谜团之中,留下了糖生物学界几十年来最大的“悬案”。本课题组致力于以上未知科学问题的研究和探索。我们的主要研究目标如下:

研究目标

1)真核细胞内质网N-糖基化修饰调控机制的阐明

  应用分子生物学、生物化学以及细胞遗传学等研究技术,针对多萜醇寡糖前体DLO合成关键糖基转移酶Alg的蛋白结构、复合体形成、反应机制以及其蛋白质降解体系等具体课题展开研究和探索,挑战“N-糖基化调控”这个糖生物学领域最关键的基础课题。

2)多萜醇寡糖翻转酶的筛选及翻转分子机制的解析

  以研究多萜醇寡糖翻转相关蛋白Rft1的确切功能为突破口,首先通过从不同真核细胞的基因文库中筛选酿酒酵母rft1温度敏感突变株的多拷贝抑制子的方法,进一步研究并确认Rft1的生理功能。同时,争取从多拷贝抑制子中发现并鉴定M5-Dol翻转酶或其他翻转体系相关蛋白编码基因,通过体外翻转酶活性测试,以及针对相应酵母或动物细胞突变菌株的进一步研究,从蛋白结构和生理活性上鉴定M5-Dol翻转酶及其相关蛋白的存在,并探索多萜醇寡糖翻转体系的分子机制。

3)真核细胞医药糖蛋白分泌表达体系的研发  

  充分利用内质网N-糖基化途径在各类真核细胞中具有高度保守性的特点,构建酵母和动物细胞外源糖蛋白表达体系,开发抗体等医药糖蛋白的生产技术。


主要科研项目 

2018-2021: 国家自然科学基金 

2013-2016:江苏省双创高层次人才引进计划

2012-2015:教育部科学技术研究重大项目 

2012-2015:高等学校博士学科点专项科研基金


2017年发表论文

SCI Articles

Corresponding author:

1) Kitajima T*, Xue W, Liu YS, Wang CD, Liu SS, Fujita M, Gao X-D* (2017) Construction of green fluorescence protein mutant to monitor STT3B-dependent N-glycosylation. FEBS J. 2017 Dec 28. doi: 10.1111/febs.14375. 

2) Li S-T, Wang N, Xu X-X, Fujita M, Nakanishi H, Kitajima T, Dean N* and Gao X-D*. (2017) Alternative routes for synthesis of N-linked glycans by the Alg2 mannosyltransferase. FASEB J. 2017 Dec 22. pii: fj.201701267R. doi: 10.1096/fj.

3) Pan HP, Wang N, Tachikawa H, Gao X-D* and Nakanishi H*. (2017) Osw2 is required for proper assembly of glucan and/or mannan layers of the yeast spore wall. J Biochem. doi: 10.1093/jb/mvx082.

4) Xu XX, Komatsuzaki A, Chiba Y, Gao X-D* and Yoko-O T*. (2017) PER1, GUP1, and CWH43 of methylotrophic yeast Ogataea minuta are involved in cell wall integrity. Yeast. 2017 Oct 13. doi: 10.1002/yea.3285. 

5) Bemena LD, Mukama O, Neiman AM, Li Z, Gao X-D* and Nakanishi H*. (2017) In vitro reconstitution of the yeast spore wall dityrosine layer to disclose the mechanism of its assembly. J Biol Chem. 2017 Sep 22;292(38):15880-15891. doi: 10.1074/jbc.

6) Pan HP, Wang N, Tachikawa H, Nakanishi H* and Gao X-D*. (2017) ß-1,6-glucan synthesis-associated genes are required for proper spore wall formation in Saccharomyces cerevisiaeYeast.2017 Jul 21. doi: 10.1002/yea.3244. doi: 10.1002/yea.3244.

7) Yan T., Zhang H.*, Huang D., Feng S., Fujita M. and Gao X-D* (2017) Chitosan-functionalized graphene oxide as potential immunoadjuvant. Nanomaterials (Basel). 2017 Mar 8;7(3). doi: 10.3390/nano7030059

8) Nakanishi H., Li F; Han B., Arai S., Gao X-D*, (2017) Yeast cells as an assay system for in vivo O-GlcNAc modification. BBA-general subjects, 2017 May; 1861(5 Pt A): 1159-1167. doi: 10.1016/j.bbagen.2017.03.002.

9) Li S-T., Wang N., Xu S., Yin J, Nakanishi H*., Dean N, Gao X-D*. (2017) Quantitative study of yeast Alg1 beta-1, 4 mannosyltransferase activity, a key enzyme involved in protein N-glycosylation. BBA-general subjects, 2017 Jan; 1861(1 Pt A): 2934-2941. doi: 10.1016/j.bbagen.2016.09.023.

10) Zhang, HJ., Yan, T., Xu, S., Feng, SN., Huang, DD., Fujita, M., and Gao X-D*(2017) Graphene oxide-chitosan nanocomposites for intracellular delivery of immunostimulatory CpG oligodeoxynucleotides. Materials Science and Engineering: C. 73:144-51 

11) Kong J., Li Z., Zhang H., Gao X-D* and Nakanishi H*. (2017) Production of encapsulated creatinase using yeast spores, Bioengineered. 2017 Jul 4;8(4):411-419. doi: 10.1080/21655979.2016.1241926. Epub 2016 Oct 28.PMID: 27791465

12) Kong J., Li Z., Zhang H., Gao X-D* and Nakanishi H*. (2017) Consecutive hydrolysis of creatinine using creatininase and creatinase encapsulated in Saccharomyces cerevisiae spores, Biotechnology Letters, 2017 Feb;39(2):261-267. DOI: 10.1007/s10529-016-2234-9. Epub 2016 Oct 12.PMID: 27734207

13) Bemena LD., Mukama O., Wang N., Gao X-D*. and Nakanishi H*. (2017) Characterization of a yeast sporulation-specific P450 family protein, Dit2, using an in vitro assay to crosslink formyl tyrosine. J Biochem. Oct 9. doi: 10.1093/jb/mvx082. 

Coauthor

14) Liu YS, Guo XY, Hirata T, Rong Y, Motooka D, Kitajima T, Murakami Y, Gao X-D, Nakamura S, Kinoshita T, Fujita M*. (2018) N-Glycan-dependent protein folding and endoplasmic reticulum retention regulate GPI-anchor processing. J Cell Biol. 2017 Dec 18. pii: jcb.201706135. doi: 10.1083/jcb.201706135.

15) Nakamura TS, Numajiri Y, Okumura Y, Hidaka J, Tanaka T, Inoue I, Suda Y, Takahashi T, Nakanishi H, Gao X-D, Neiman AM, Tachikawa H*. (2017) Dynamic localization of a yeast development-specific PP1 complex during prospore membrane formation is dependent on multiple localization signals and complex formation. Mol Biol Cell. 2017 Oct 18. pii: mbc. E17-08-0521. doi: 10.1091/mbc.

16) Kitajima T, Jia Y, Komatsuzaki A, Cui J, Matsuzawa F, Aikawa SI, Gao X-D, Chiba Y*. (2017) Structural modeling and mutagenesis of endo-β-N-acetylglucosaminidase from Ogataea minuta identifies the importance of Trp295 for hydrolytic activity. J Biosci Bioeng. 2017 Sep 11. pii: S1389-1723(17)30571-6.

  17)  Matabaro E., He Z’a., Liu Y-S., Zhang H., Gao X-D and Fujita M*. (2017) Molecular switching system using glycosylphosphatidylinositol to select cells highly expressing recombinant proteins. Scientific Rep. Jun 22; 7(1): 4033.Doi: 10.1038/s41598-017-04330-3.

Reviews & Books

1) Li A., Cai L., Chen Z., Wang M., Wang N., Nakanishi H, Gao X-D*, Zijie Li,* (2017) Recent advances in the synthesis of rare sugars using DHAP-Dependent aldolases. Carbohydrate Research · October 2017. DOI: 10.1016/j.carres.2017.10.009

2) Wang N., Li S-T., Lu T., Nakanishi H, Gao X-D*. (2017) Approaches towards the core pentasaccharide in N-linked glycans. Chinese Chemical Letters, 2017, doi 10.1016/j.cclet.2017.09.044

3) Zhang H and Gao X-D*(2017) Nanodelivery systems for enhancing the immunostimulatory effect of CpG oligodeoxynucleotides. Mater Sci Eng C Mater Biol Appl. 2017 Jan 1; 70(Pt 2):935-946. doi: 10.1016/j.msec.2016.03.045.  


代表性科研成果  

SCI论文  

1) Li S-T, Wang N, Xu X-X, Fujita M, Nakanishi H, Kitajima T, Dean N* and Gao X-D*. (2017) Alternative routes for synthesis of N-linked glycans by the Alg2 mannosyltransferase. FASEB J. 2017 Dec 22. pii: fj.201701267R. doi: 10.1096/fj.

2) Kitajima T*, Xue W, Liu YS, Wang CD, Liu SS, Fujita M, Gao X-D* (2017) Construction of green fluorescence protein mutant to monitor STT3B-dependent N-glycosylation. FEBS J. 2017 Dec 28. doi: 10.1111/febs.14375. 

3) Bemena LD, Mukama O, Neiman AM, Li Z, Gao X-D* and Nakanishi H*.In vitro reconstitution of the yeast spore wall dityrosine layer to disclose the mechanism of its assembly. J Biol Chem. 2017 Sep 22;292(38):15880-15891. doi: 10.1074/jbc.

4) Li S-T., Wang N., Xu S., Yin J, Nakanishi H*., Dean N, Gao X-D*, (2017) Quantitative study of yeast Alg1 beta-1, 4 mannosyltransferase activity, a key enzyme involved in protein N-glycosylation. BBA-general subjects, 2017 Jan; 1861(1 Pt A): 2934-2941 

5) Nakanishi H., Li F; Han B., Arai S., Gao X-D*, (2017) Yeast cells as an assay system for in vivo O-GlcNAc modification. BBA-general subjects, 2017 May; 1861(5 Pt A): 1159-1167. doi: 10.1016/j.bbagen.2017.03.002.

6) Gao X-D, Moriyama S, Miura N, Dean N and Nishimura S-I*. (2008) Interaction between the C-termini of Alg13 and Alg14 mediate formation of the active UDP-N-acetylglucosamine transferase complex. J. Biol. Chem., 283: 32534-32541  

7) Gao X-D, TachikawaH, SatoT, JigamiY and Dean N*. (2005) Alg14 recruits Alg13 to the cytoplasmic face of the endoplasmic reticulum to form a novel bipartite UDP-N-acetylglucosaminetransferase required for the second step of N-linked glycosylation. J. Biol. Chem., 280: 36254 – 36262  

8) Gao X-D, Nishikawa A, and Dean N*. (2001) Identification of a conserved motif in the yeast GDP-mannose transporter required for nucleotide sugar binding. J. Biol. Chem., 276: 4424-4432.  

9) Gao X-D and Dean, N*. (2000) Distinct protein domains of the yeast Golgi GDP-mannose transporter mediate oligomer assembly and export from the endoplasmic reticulum. J. Biol. Chem., 275: 17718-17727.  

10) Gao X-D, Kaigorodov V and Jigami Y*. (1999) YND1, a homologue of GDA1, encodes membrane bound apyrase required for Golgi N- and 0-glycosylation in Saccharomyes cerevisiae., J. Biol. Chem., 274: 21450 – 21456. 

Reviews & Books 

1) Zhang H and Gao X-D*. Nanodelivery systems for enhancing the immunostimulatory effect of CpG oligodeoxynucleotides. Mater Sci Eng C Mater Biol Appl. 2017 Jan 1; 70(Pt 2):935-946. doi: 10.1016/j.msec.2016.03.045. (2017)

2) Wang N., Li S-T., Lu T., Nakanishi H, Gao X-D*. (2017) Approaches towards the core pentasaccharide in N-linked glycans. Chinese Chemical Letters, 2017, doi 10.1016/j.cclet.2017.09.044

3) Dean N, Gao X-D*.: Heterodimeric Alg13/Alg14 UDP-GlcNAc transferase (ALG13,14). In: Taniguchi N., Honke K., Fukuda M., Narimatsu H., Yamaguchi Y., Angata T. (Ed.) Handbook of Glycosyltransferases and Related Genes: Springer-Verlag Berlin Heidelberg, DOI: 10.1007/SpringerReference_332094 2013-05-26 14:16:42 UTC, (2013)  

4) Dean N, Gao X-D*.: Dolichyl-phosphate (UDP-N-acetylglucosamine) N-acetylglucosaminephosphotransferase 1 (GlcNAc-1-P transferase) (DPAGT1). In: Taniguchi N., Honke K., Fukuda M., Narimatsu H., Yamaguchi Y., Angata T. (Ed.) Handbook of Glycosyltransferases and Related Genes:. Springer-Verlag Berlin Heidelberg, DOI: 10.1007/SpringerReference_332181 2013-05-26 14:04:03 UTC,(2013)


所获专利
1)地神 芳文、千木良 裕子、高 暁冬: O-フコース結合タンパク質合成系遺伝子が導入された酵母形質転換体、特許第4258662号(JAPAN)、February 20th.2009
2)中西秀树,高晓冬,张海妮,李子杰.一种以酿酒酵母孢子为载体的新型固定化酶的制备方法. 2013.9,中国,专利号:201310422568.8

 

荣誉&奖励
1)2013年:    “江苏省六大人才高峰”
2)2011年: “江苏省双创高层次人才引进计划”
3)2001年:  Selected and Interviewed as “Dedicated postdoc” by AAAS  [Ref: Science, 294(5541): p410 (2001)]
4)2000年: 荣获美国杰出科技人才签证(O1) 
5)1984年: 中国教育部派遣本科留学生(April 1984 --- March 1993)

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