Role of the human ST6GalNAc-I and ST6GalNAc-II in the synthesis of the cancer-associated Sialyl-Tn antigen

Nuno T. Marcos, Sandra I. Pinho, Catarina Grandela, Andrea Cruz, Bénédicte Samyn-Petit, Anne Harduin-Lepers, Raquel Almeida, Filipe Silva, Vanessa Morais, Julia Costa, Jan Kihlberg, Henrik Clausen, Celso A. Reis

Research output: Contribution to journalArticle

131 Citations (Scopus)

Abstract

The Sialyl-Tn antigen (Neu5Acα2-6GalNAc-O-Ser/Thr) is highly expressed in several human carcinomas and is associated with carcinoma aggressiveness and poor prognosis. We characterized two human sialyltransferases, CMP-Neu5Ac:GalNAc-R α2,0-sialyltransferase (ST6GalNAc)-I and ST6GalNAc-II, that are candidate enzymes for Sialyl-Tn synthases. We expressed soluble recombinant hST6GalNAc-I and hST6GalNAc-II and characterized the substrate specificity of both enzymes toward a panel of glycopeptides, glycoproteins, and other synthetic glycoconjugates. The recombinant ST6GalNAc-I and ST6GalNAc-II showed similar substrate specificity toward glycoproteins and GalNAcα-O-Ser/Thr glycopeptides, such as glycopeptides derived from the MUC2 mucin and the HIVgp120. We also observed that the amino acid sequence of the acceptor glycopeptide contributes to the in vitro substrate specificity of both enzymes. We additionally established a gastric cell line, MKN45, stably transfected with the full length of either ST6GalNAc-I or ST6GalNAc-II and evaluated the carbohydrate antigens expression profile induced by each enzyme. MKN45 transfected with ST6GalNAc-I showed high expression of Sialyl-Tn, whereas MKN45 transfected with ST6GalNAc-II showed the biosynthesis of the Sialyl-6T structure [Galβ1-3 (Neu5Acα2-6)GalNAc- O-Ser/Thr]. In conclusion, although both enzymes show similar in vitro activities when Tn antigen alone is available, whenever both Tn and T antigens are present, ST6GalNAc-I acts preferentially on Tn antigen, whereas the ST6GalNAc-II acts preferentially on T antigen. Our results show that ST6GalNAc-I is the major Sialyl-Tn synthase and strongly support the hypothesis that the expression of the Sialyl-Tn antigen in cancer cells is due to ST6GalNAc-I activity.

Original languageEnglish (US)
Pages (from-to)7050-7057
Number of pages8
JournalCancer Research
Volume64
Issue number19
DOIs
StatePublished - Oct 1 2004
Externally publishedYes

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Glycopeptides
Substrate Specificity
Sialyltransferases
Enzymes
Viral Tumor Antigens
Neoplasms
Glycoproteins
Carcinoma
Glycoconjugates
Mucins
Amino Acid Sequence
Stomach
Carbohydrates
galactosyl-1-3-N-acetylgalactosaminyl-specific 2,6-sialyltransferase
sialosyl-Tn antigen
Antigens
Cell Line
Tn antigen
In Vitro Techniques

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Marcos, N. T., Pinho, S. I., Grandela, C., Cruz, A., Samyn-Petit, B., Harduin-Lepers, A., ... Reis, C. A. (2004). Role of the human ST6GalNAc-I and ST6GalNAc-II in the synthesis of the cancer-associated Sialyl-Tn antigen. Cancer Research, 64(19), 7050-7057. https://doi.org/10.1158/0008-5472.CAN-04-1921

Role of the human ST6GalNAc-I and ST6GalNAc-II in the synthesis of the cancer-associated Sialyl-Tn antigen. / Marcos, Nuno T.; Pinho, Sandra I.; Grandela, Catarina; Cruz, Andrea; Samyn-Petit, Bénédicte; Harduin-Lepers, Anne; Almeida, Raquel; Silva, Filipe; Morais, Vanessa; Costa, Julia; Kihlberg, Jan; Clausen, Henrik; Reis, Celso A.

In: Cancer Research, Vol. 64, No. 19, 01.10.2004, p. 7050-7057.

Research output: Contribution to journalArticle

Marcos, NT, Pinho, SI, Grandela, C, Cruz, A, Samyn-Petit, B, Harduin-Lepers, A, Almeida, R, Silva, F, Morais, V, Costa, J, Kihlberg, J, Clausen, H & Reis, CA 2004, 'Role of the human ST6GalNAc-I and ST6GalNAc-II in the synthesis of the cancer-associated Sialyl-Tn antigen', Cancer Research, vol. 64, no. 19, pp. 7050-7057. https://doi.org/10.1158/0008-5472.CAN-04-1921
Marcos NT, Pinho SI, Grandela C, Cruz A, Samyn-Petit B, Harduin-Lepers A et al. Role of the human ST6GalNAc-I and ST6GalNAc-II in the synthesis of the cancer-associated Sialyl-Tn antigen. Cancer Research. 2004 Oct 1;64(19):7050-7057. https://doi.org/10.1158/0008-5472.CAN-04-1921
Marcos, Nuno T. ; Pinho, Sandra I. ; Grandela, Catarina ; Cruz, Andrea ; Samyn-Petit, Bénédicte ; Harduin-Lepers, Anne ; Almeida, Raquel ; Silva, Filipe ; Morais, Vanessa ; Costa, Julia ; Kihlberg, Jan ; Clausen, Henrik ; Reis, Celso A. / Role of the human ST6GalNAc-I and ST6GalNAc-II in the synthesis of the cancer-associated Sialyl-Tn antigen. In: Cancer Research. 2004 ; Vol. 64, No. 19. pp. 7050-7057.
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abstract = "The Sialyl-Tn antigen (Neu5Acα2-6GalNAc-O-Ser/Thr) is highly expressed in several human carcinomas and is associated with carcinoma aggressiveness and poor prognosis. We characterized two human sialyltransferases, CMP-Neu5Ac:GalNAc-R α2,0-sialyltransferase (ST6GalNAc)-I and ST6GalNAc-II, that are candidate enzymes for Sialyl-Tn synthases. We expressed soluble recombinant hST6GalNAc-I and hST6GalNAc-II and characterized the substrate specificity of both enzymes toward a panel of glycopeptides, glycoproteins, and other synthetic glycoconjugates. The recombinant ST6GalNAc-I and ST6GalNAc-II showed similar substrate specificity toward glycoproteins and GalNAcα-O-Ser/Thr glycopeptides, such as glycopeptides derived from the MUC2 mucin and the HIVgp120. We also observed that the amino acid sequence of the acceptor glycopeptide contributes to the in vitro substrate specificity of both enzymes. We additionally established a gastric cell line, MKN45, stably transfected with the full length of either ST6GalNAc-I or ST6GalNAc-II and evaluated the carbohydrate antigens expression profile induced by each enzyme. MKN45 transfected with ST6GalNAc-I showed high expression of Sialyl-Tn, whereas MKN45 transfected with ST6GalNAc-II showed the biosynthesis of the Sialyl-6T structure [Galβ1-3 (Neu5Acα2-6)GalNAc- O-Ser/Thr]. In conclusion, although both enzymes show similar in vitro activities when Tn antigen alone is available, whenever both Tn and T antigens are present, ST6GalNAc-I acts preferentially on Tn antigen, whereas the ST6GalNAc-II acts preferentially on T antigen. Our results show that ST6GalNAc-I is the major Sialyl-Tn synthase and strongly support the hypothesis that the expression of the Sialyl-Tn antigen in cancer cells is due to ST6GalNAc-I activity.",
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T1 - Role of the human ST6GalNAc-I and ST6GalNAc-II in the synthesis of the cancer-associated Sialyl-Tn antigen

AU - Marcos, Nuno T.

AU - Pinho, Sandra I.

AU - Grandela, Catarina

AU - Cruz, Andrea

AU - Samyn-Petit, Bénédicte

AU - Harduin-Lepers, Anne

AU - Almeida, Raquel

AU - Silva, Filipe

AU - Morais, Vanessa

AU - Costa, Julia

AU - Kihlberg, Jan

AU - Clausen, Henrik

AU - Reis, Celso A.

PY - 2004/10/1

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N2 - The Sialyl-Tn antigen (Neu5Acα2-6GalNAc-O-Ser/Thr) is highly expressed in several human carcinomas and is associated with carcinoma aggressiveness and poor prognosis. We characterized two human sialyltransferases, CMP-Neu5Ac:GalNAc-R α2,0-sialyltransferase (ST6GalNAc)-I and ST6GalNAc-II, that are candidate enzymes for Sialyl-Tn synthases. We expressed soluble recombinant hST6GalNAc-I and hST6GalNAc-II and characterized the substrate specificity of both enzymes toward a panel of glycopeptides, glycoproteins, and other synthetic glycoconjugates. The recombinant ST6GalNAc-I and ST6GalNAc-II showed similar substrate specificity toward glycoproteins and GalNAcα-O-Ser/Thr glycopeptides, such as glycopeptides derived from the MUC2 mucin and the HIVgp120. We also observed that the amino acid sequence of the acceptor glycopeptide contributes to the in vitro substrate specificity of both enzymes. We additionally established a gastric cell line, MKN45, stably transfected with the full length of either ST6GalNAc-I or ST6GalNAc-II and evaluated the carbohydrate antigens expression profile induced by each enzyme. MKN45 transfected with ST6GalNAc-I showed high expression of Sialyl-Tn, whereas MKN45 transfected with ST6GalNAc-II showed the biosynthesis of the Sialyl-6T structure [Galβ1-3 (Neu5Acα2-6)GalNAc- O-Ser/Thr]. In conclusion, although both enzymes show similar in vitro activities when Tn antigen alone is available, whenever both Tn and T antigens are present, ST6GalNAc-I acts preferentially on Tn antigen, whereas the ST6GalNAc-II acts preferentially on T antigen. Our results show that ST6GalNAc-I is the major Sialyl-Tn synthase and strongly support the hypothesis that the expression of the Sialyl-Tn antigen in cancer cells is due to ST6GalNAc-I activity.

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