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Promotion of cell migration by neural cell adhesion molecule (NCAM) is enhanced by PSA in a polysialyltransferase-specific manner.

Guan F, Wang X, He F - PLoS ONE (2015)

Bottom Line: We cloned the genes for NCAM-140 and for polysialyltransferases STX and PST (responsible for PSA synthesis) from normal murine mammary gland epithelial (NMuMG) cells and transfected them into ldlD-14 and human breast cancer cells MCF-7.The effect of PSA on NCAM-mediated cell proliferation, motility, migration and adhesion was studied.We found that NCAM-140 significantly promoted cell proliferation, motility and migration, while polysialylation of NCAM-140 catalyzed by STX, but not by PST, enhanced NCAM-mediated cell migration, but not cell proliferation or motility.

View Article: PubMed Central - PubMed

Affiliation: The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China.

ABSTRACT
Neural cell adhesion molecule 140 (NCAM-140) is a glycoprotein and always highly polysialylated in cancer. Functions of polysialic acid (PSA) that binds to N-glycan termini on NCAM remain unclear. ldlD-14 cells, a CHO cell mutant deficient in UDP-Gal 4-epimerase, are useful for structural and functional studies of Gal-containing glycoproteins because their abnormal glycosylation can be converted to normal status by exogenous addition of galactose (Gal). We cloned the genes for NCAM-140 and for polysialyltransferases STX and PST (responsible for PSA synthesis) from normal murine mammary gland epithelial (NMuMG) cells and transfected them into ldlD-14 and human breast cancer cells MCF-7. The effect of PSA on NCAM-mediated cell proliferation, motility, migration and adhesion was studied. We found that NCAM-140 significantly promoted cell proliferation, motility and migration, while polysialylation of NCAM-140 catalyzed by STX, but not by PST, enhanced NCAM-mediated cell migration, but not cell proliferation or motility. In addition, PSA catalyzed by different polysialyltransferases affected the adhesion of NCAM to different extracellular matrix (ECM) components.

No MeSH data available.


Related in: MedlinePlus

NCAM and PSA expression (schematic) in ldlD/N140 cells cultured in ITS with or without Gal.(A) Structure of NCAM-140. The extracellular domain (ECD) of NCAM is composed of five immunoglobulin (Ig)-like domains and two fibronectin type III (F3) repeats. NCAM has six N-glycosylation sites, of which the 5th and 6th have N-glycans that are modified by sialic acid or PSA. TMD, transmembrane domain; ICD, intercellular domain. (B) In cells cultured with ITS alone, NCAM-140 has no PSA modification due to the lack of Gal on terminal of deficient N-glycans. (C) In cells cultured with ITS+Gal, PSA-NCAM is attached on the NCAM. In the structural diagram, n≥1.
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pone.0124237.g007: NCAM and PSA expression (schematic) in ldlD/N140 cells cultured in ITS with or without Gal.(A) Structure of NCAM-140. The extracellular domain (ECD) of NCAM is composed of five immunoglobulin (Ig)-like domains and two fibronectin type III (F3) repeats. NCAM has six N-glycosylation sites, of which the 5th and 6th have N-glycans that are modified by sialic acid or PSA. TMD, transmembrane domain; ICD, intercellular domain. (B) In cells cultured with ITS alone, NCAM-140 has no PSA modification due to the lack of Gal on terminal of deficient N-glycans. (C) In cells cultured with ITS+Gal, PSA-NCAM is attached on the NCAM. In the structural diagram, n≥1.

Mentions: We used an ldlD-14 cell model to elucidate the role of PSA in NCAM function. The UDP-Gal 4-epimerase deficiency characteristic of ldlD-14 cells results in low internal pools of UDP-Gal when cells are grown in the absence of Gal [33]. The derived cell line ldlD-14 is a useful model for studying the functions of glycans in glycoproteins and glycolipids[18,42]. NCAM model in ldlD-14 cells was proposed when cultured in ITS only condition. In this situation, incomplete N-glycan on ldlD-14 cells lacked the factors of Gal for the attachment of PSA (Fig 7B). To study the role of PSA-NCAM, addition of Gal on N-glycan termini is the essential step for PSA synthesis on NCAM (Fig 7C). Using transfected ldlD/N140 cells, we were able to restore normal cell phenotype by addition of Gal and study glycan patterns by flow cytometry and MALDI-TOF/TOF-MS/MS.


Promotion of cell migration by neural cell adhesion molecule (NCAM) is enhanced by PSA in a polysialyltransferase-specific manner.

Guan F, Wang X, He F - PLoS ONE (2015)

NCAM and PSA expression (schematic) in ldlD/N140 cells cultured in ITS with or without Gal.(A) Structure of NCAM-140. The extracellular domain (ECD) of NCAM is composed of five immunoglobulin (Ig)-like domains and two fibronectin type III (F3) repeats. NCAM has six N-glycosylation sites, of which the 5th and 6th have N-glycans that are modified by sialic acid or PSA. TMD, transmembrane domain; ICD, intercellular domain. (B) In cells cultured with ITS alone, NCAM-140 has no PSA modification due to the lack of Gal on terminal of deficient N-glycans. (C) In cells cultured with ITS+Gal, PSA-NCAM is attached on the NCAM. In the structural diagram, n≥1.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4401701&req=5

pone.0124237.g007: NCAM and PSA expression (schematic) in ldlD/N140 cells cultured in ITS with or without Gal.(A) Structure of NCAM-140. The extracellular domain (ECD) of NCAM is composed of five immunoglobulin (Ig)-like domains and two fibronectin type III (F3) repeats. NCAM has six N-glycosylation sites, of which the 5th and 6th have N-glycans that are modified by sialic acid or PSA. TMD, transmembrane domain; ICD, intercellular domain. (B) In cells cultured with ITS alone, NCAM-140 has no PSA modification due to the lack of Gal on terminal of deficient N-glycans. (C) In cells cultured with ITS+Gal, PSA-NCAM is attached on the NCAM. In the structural diagram, n≥1.
Mentions: We used an ldlD-14 cell model to elucidate the role of PSA in NCAM function. The UDP-Gal 4-epimerase deficiency characteristic of ldlD-14 cells results in low internal pools of UDP-Gal when cells are grown in the absence of Gal [33]. The derived cell line ldlD-14 is a useful model for studying the functions of glycans in glycoproteins and glycolipids[18,42]. NCAM model in ldlD-14 cells was proposed when cultured in ITS only condition. In this situation, incomplete N-glycan on ldlD-14 cells lacked the factors of Gal for the attachment of PSA (Fig 7B). To study the role of PSA-NCAM, addition of Gal on N-glycan termini is the essential step for PSA synthesis on NCAM (Fig 7C). Using transfected ldlD/N140 cells, we were able to restore normal cell phenotype by addition of Gal and study glycan patterns by flow cytometry and MALDI-TOF/TOF-MS/MS.

Bottom Line: We cloned the genes for NCAM-140 and for polysialyltransferases STX and PST (responsible for PSA synthesis) from normal murine mammary gland epithelial (NMuMG) cells and transfected them into ldlD-14 and human breast cancer cells MCF-7.The effect of PSA on NCAM-mediated cell proliferation, motility, migration and adhesion was studied.We found that NCAM-140 significantly promoted cell proliferation, motility and migration, while polysialylation of NCAM-140 catalyzed by STX, but not by PST, enhanced NCAM-mediated cell migration, but not cell proliferation or motility.

View Article: PubMed Central - PubMed

Affiliation: The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China.

ABSTRACT
Neural cell adhesion molecule 140 (NCAM-140) is a glycoprotein and always highly polysialylated in cancer. Functions of polysialic acid (PSA) that binds to N-glycan termini on NCAM remain unclear. ldlD-14 cells, a CHO cell mutant deficient in UDP-Gal 4-epimerase, are useful for structural and functional studies of Gal-containing glycoproteins because their abnormal glycosylation can be converted to normal status by exogenous addition of galactose (Gal). We cloned the genes for NCAM-140 and for polysialyltransferases STX and PST (responsible for PSA synthesis) from normal murine mammary gland epithelial (NMuMG) cells and transfected them into ldlD-14 and human breast cancer cells MCF-7. The effect of PSA on NCAM-mediated cell proliferation, motility, migration and adhesion was studied. We found that NCAM-140 significantly promoted cell proliferation, motility and migration, while polysialylation of NCAM-140 catalyzed by STX, but not by PST, enhanced NCAM-mediated cell migration, but not cell proliferation or motility. In addition, PSA catalyzed by different polysialyltransferases affected the adhesion of NCAM to different extracellular matrix (ECM) components.

No MeSH data available.


Related in: MedlinePlus