人類岩藻醣水解酶之生化性質探討

高敏絹

3.239.33.139，Hello！

Close
Search Quick Search Advanced Search Publication Search	Personal Service Member Login Searching History Use Bonus Card SYMSKAN
Browse Browse by University of Graduate Thesis Browse by Pinyin of Graduate Thesis Browse by Conference Proceedings Topic Browse by Conference Proceedings Publisher Browse by Pinyin of Conference Proceedings Browse by eBook Category Browse by eBook Topic Browse by eBook Publisher
Customer Services Authorize Airiti Recommend a Journal

Search Symbol (Half-width)	Description of Search Symbols
Space	"AND" indicates the intertwining of key terms used in a search
Double Quotation Marks ("") ( " " )	Double quotation marks indicate the beginning and end of a phrase, and the search will only include terms that appear in the same order of those within the quotations. Example: "image process" ： " image process "
?	Indicates a variable letter. Entering two ? will indicate two variable letters, and so on. Example: "Appl?", search results will yield apple, apply… e , appl y … ( (often used to English word searches) )
*	Indicates an unlimited number of variable letters to follow, from 1~n. Example: Enter "appl*", search results will yield apple, apples, apply, applied, application…(often used in English word searches) e , appl es , appl y , appl ied , appl ication … ( (often used to English word searches) )
AND、OR、NOT	Boolean logic combinations of key words is a skill used to expand or refine search parameters. (1) AND (1) AND: Refines search parameters (2) OR (2) OR: Expands search parameters (3) NOT: Excludes irrelevant parameters

DOI stands for Digital Object Identifier （ D igital O bject I dentifier ） ,
and is the unique identifier for objects on the internet. It can be used to create persistent link and to cite articles.

Using DOI as a persistent link

To create a persistent link, add「http://dx.doi.org/」「 http://dx.doi.org/ 」 before a DOI.
For instance, if the DOI of an article is 10.5297/ser.1201.002 , you can link persistently to the article by entering the following link in your browser: http://dx.doi.org/ 10.5297/ser.1201.002 。
The DOI link will always direct you to the most updated article page no matter how the publisher changes the document's position, avoiding errors when engaging in important research.

Cite a document with DOI

When citing references, you should also cite the DOI if the article has one. If your citation guideline does not include DOIs, you may cite the DOI link.

DOIs allow accurate citations, improve academic contents connections, and allow users to gain better experience across different platforms. Currently, there are more than 70 million DOIs registered for academic contents. If you want to understand more about DOI, please visit airiti DOI Registration （ doi.airiti.com ）。

Data Source

College of Life Science > Graduate Institute of Biochemical Sciences
Bio-Agriculture > Biological Science

Bibliography Management Tool

Related Links

Report an Issue

Purchase Single Articles

Download PDF

人類岩藻醣水解酶之生化性質探討

Characterization of Human α-L-Fucosidase: Large-scale preparation, substrate specificity

高敏絹 , Masters　　Advisor：林俊宏　　

繁體中文

岩藻醣水解酶受質專一性；肝癌；腫瘤指標； α-L-fucosidases ； substrate specificity ； liver cancer ； biomarker

Abstract │ Reference (113) │ Altmetrics

Abstract 〈TOP〉

Parallel Abstract 〈TOP〉

There are two α-L-fucosidases (EC 3.2.1.51) in human, including α-L-fucosidase1 (FucA1) and α-L-fucosidase2 (FucA2). They are responsible for the removal of L-fucose residues from the non-reducing end of glycoconjugates. Abnormal fucosidase activity has been associated with many diseases, such as hepatocellular cancer and breast cancer. Previous study indicated that the infection of Helicobacter pylori triggers the release of FucA2 by gastric cancer cells. However, the role and physiological function of FucA2 still remain ambiguous. One of the prerequisites for further studies is to prepare FucA2 in a sufficient amount and to carry out their characterizations.
     Previous preparation of FucA2 in our labotoary was restricted by the poor expression and extremely low activity. Herein we succesfully express recombinant FucA2 in E. coli.by incorporating 6x His, MBP and TEV protease recognition site to N-terminus of FucA2. Significant expression and activity were obtained after removal of the MBP-fusion tag with TEV protease, allowing the biochemical characterizations such as pH profile, kinetic parameters and substrate specificity. The results indicated that FucA2 is optimally active at pH 6.0, unlike FucA1 which has dual optimum at pH 4.5 and pH 6.5.
     To examine the substrate specificity, the glycoproteins extracted from human colon cancer cells line (Colo 205) and seven pyridylamino-conjugated sugars were used for the study. These glycoconjugates were subjected to the hydrolytic cleavage of either FucA1 or FucA2, followed by mass spectrometric and HPLC analyse, respectively the results indicate that FucA1 preferentially hydrolyzes L-fucose from Fuc α-1,2 Gal β-1,4, core Fuc α-1,6 bi/triantennary, Lea, Lex, while FUCA2 catalyzes the hydrolysis of Fuc α-1,2 Gal β-1,4, and Lex.
     Furthermore, both recombinant FucA1 and FucA2 were examined by the antibodies that were generated by immunization of the specific peptides correspounding to residues 404-444 of FucA1 and residues 404-445 of FucA2 (represent the major sequence differance). In conjunction with our developed inhibitor-based affinity column chromatography, the antibodies were used to examine and identify the serum fucosidase from the patients with liver and gastric cancers. The result indicated that high activity of FucA1 was found in the serum of liver cancer patients, while that of FucA2 in the serum of normal people. Further examination (such as the glycol-structure analysis) would be beneficial to use FucA1 as a biomarker.

Reference ( 113 ) 〈TOP〉

1 Delves, P. J. The role of glycosylation in autoimmune disease. Autoimmunity 27, 239-253,(1998).
2 Moloney, D. J., Shair, L. H., Lu, F. M., Xia, J., Locke, R., Matta, K. L. & Haltiwanger, R. S. Mammalian Notch1 is modified with two unusual forms of O-linked glycosylation found on epidermal growth factor-like modules. J Biol Chem 275, 9604-9611,(2000).
3 Haltiwanger, R. S. & Lowe, J. B. Role of glycosylation in development. Annu Rev Biochem 73, 491-537,(2004).
4 Greenwell, P. Blood group antigens: molecules seeking a function? Glycoconj J 14, 159-173,(1997).
5 Ma, B., Simala-Grant, J. L. & Taylor, D. E. Fucosylation in prokaryotes and eukaryotes. Glycobiology 16, 158R-184R,(2006).

Altmetrics

〈TOP〉

E-mail ：

When an article is available to download, a notice will be sent to your mailbox address.

E-mail ：