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Expression of (alpha1,3) fucosyltran...

Chien, Wenwen,

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Expression of (alpha1,3) fucosyltransferase in B16 mouse melanoma cells alters cell adhesive properties /

Record Type:	Electronic resources : Monograph/item
Title/Author:	Expression of (alpha1,3) fucosyltransferase in B16 mouse melanoma cells alters cell adhesive properties // Wenwen Chien.
Author:	Chien, Wenwen,
Description:	1 electronic resource (137 pages)
Notes:	Source: Dissertations Abstracts International, Volume: 60-05, Section: B.
Contained By:	Dissertations Abstracts International60-05B.
Subject:	Cellular biology. -
Online resource:	https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=9828372
ISBN:	9780591809329

Expression of (alpha1,3) fucosyltransferase in B16 mouse melanoma cells alters cell adhesive properties /
Chien, Wenwen,

Expression of (alpha1,3) fucosyltransferase in B16 mouse melanoma cells alters cell adhesive properties /Wenwen Chien. - 1 electronic resource (137 pages)

Source: Dissertations Abstracts International, Volume: 60-05, Section: B.

Metastatic mouse melanoma B16 cells and their weakly metastatic wheat germ agglutinin-resistant Wa4, differ in the expression of an $\\alpha$1,3 fucosyltransferase activity, and this difference has been suggested to be responsible for the changes in cell adhesion and metastatic potential observed in the Wa4 cell line. To test the hypothesis that cell adhesion can be regulated solely by $\\alpha$1,3 fucosylation, parental B16 cells were stably transfected with a cDNA encoding an $\\alpha$1,3 fucosyltransferase. Clones were selected and screened for a Wa4-like phenotype. One clone, FT4, showed LTA agglutination and $\\alpha$1,3 fucosyltransferase activity similar to that of the Wa4 cells. Serial lectin affinity chromatography was used to characterize the N-linked oligosaccharides in B16, Wa4, and FT4 cells. Concanavalin A affinity chromatography suggested that the three cell lines had very similar ratios of N-linked oligosaccharides of bi-, tri-, and tetraanternary complex-type and high mannose type. Anion exchange chromatography and LTA affinity chromatography showed that FT4 and Wa4 had reduced amounts of sialic acid compared to parental B16 cells, and both contained Lewis X (Gal- ($\\alpha$1,3-Fuc) -GlcNAc) and sialyl Lewis X (NeuNAc-$\\alpha$2,3-Gal- ($\\alpha$1,3-Fuc) -GlcNAc) structures. The presence of Lewis X and sialyl Lewis X structures were further confirmed by immunofluorescence microscopy and flow cytometry using antibodies against these two structures. Analysis of proteins by lectin- and immunoblots revealed glycosylation differences including the presence of fucosylated glycoproteins in FT4 and Wa4, but not in B16. When B16, Wa4, and FT4 cells were tested for their adhesion to fibronectin and laminin, Wa4 cells and FT4 cells were both found to adhere and flatten less well than B16 cells. Homotypic adhesion assay showed increases in Wa4 and FT4 cells. These results demonstrate that a high level of expression of an endogenous or transfected $\\alpha$1,3 fucosyltransferase in B16 cells directs the synthesis of $\\alpha$1,3 fucosylated oligosaccharides, and that these changes in glycosylation were sufficient to cause a decrease in the adhesion of the cells to laminin and increase in homotypic adhesion.

English

ISBN: 9780591809329Subjects--Topical Terms:

3172791
Cellular biology.
Subjects--Index Terms:

Wa4

Expression of (alpha1,3) fucosyltransferase in B16 mouse melanoma cells alters cell adhesive properties /
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245 1 0 $a Expression of (alpha1,3) fucosyltransferase in B16 mouse melanoma cells alters cell adhesive properties / $c Wenwen Chien.
264 1 $a Ann Arbor : $b ProQuest Dissertations & Theses, $c 1998
300 $a 1 electronic resource (137 pages)
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500 $a Source: Dissertations Abstracts International, Volume: 60-05, Section: B.
500 $a Publisher info.: Dissertation/Thesis.
500 $a Advisors: Pierce, James Michael.
502 $b Ph.D. $c University of Georgia $d 1998.
520 $a Metastatic mouse melanoma B16 cells and their weakly metastatic wheat germ agglutinin-resistant Wa4, differ in the expression of an $\\alpha$1,3 fucosyltransferase activity, and this difference has been suggested to be responsible for the changes in cell adhesion and metastatic potential observed in the Wa4 cell line. To test the hypothesis that cell adhesion can be regulated solely by $\\alpha$1,3 fucosylation, parental B16 cells were stably transfected with a cDNA encoding an $\\alpha$1,3 fucosyltransferase. Clones were selected and screened for a Wa4-like phenotype. One clone, FT4, showed LTA agglutination and $\\alpha$1,3 fucosyltransferase activity similar to that of the Wa4 cells. Serial lectin affinity chromatography was used to characterize the N-linked oligosaccharides in B16, Wa4, and FT4 cells. Concanavalin A affinity chromatography suggested that the three cell lines had very similar ratios of N-linked oligosaccharides of bi-, tri-, and tetraanternary complex-type and high mannose type. Anion exchange chromatography and LTA affinity chromatography showed that FT4 and Wa4 had reduced amounts of sialic acid compared to parental B16 cells, and both contained Lewis X (Gal- ($\\alpha$1,3-Fuc) -GlcNAc) and sialyl Lewis X (NeuNAc-$\\alpha$2,3-Gal- ($\\alpha$1,3-Fuc) -GlcNAc) structures. The presence of Lewis X and sialyl Lewis X structures were further confirmed by immunofluorescence microscopy and flow cytometry using antibodies against these two structures. Analysis of proteins by lectin- and immunoblots revealed glycosylation differences including the presence of fucosylated glycoproteins in FT4 and Wa4, but not in B16. When B16, Wa4, and FT4 cells were tested for their adhesion to fibronectin and laminin, Wa4 cells and FT4 cells were both found to adhere and flatten less well than B16 cells. Homotypic adhesion assay showed increases in Wa4 and FT4 cells. These results demonstrate that a high level of expression of an endogenous or transfected $\\alpha$1,3 fucosyltransferase in B16 cells directs the synthesis of $\\alpha$1,3 fucosylated oligosaccharides, and that these changes in glycosylation were sufficient to cause a decrease in the adhesion of the cells to laminin and increase in homotypic adhesion.
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650 4 $a Cellular biology. $3 3172791
650 4 $a Oncology. $3 751006
653 $a Wa4
653 $a alpha 1,3 fucosyltransferase
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690 $a 0992
710 2 $a University of Georgia. $e degree granting institution. $3 3818195
720 1 $a Pierce, James Michael $e degree supervisor.
773 0 $t Dissertations Abstracts International $g 60-05B.
790 $a 0077
791 $a Ph.D.
792 $a 1998
856 4 0 $u https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=9828372