Human ovarian cancer, lymphoma spleen, and bovine milk GlcNAc:β1,4Gal/GalNAc transferases: Two molecular species in ovarian tumor and induction of GalNAcβ1,4Glc synthesis by α-lactalbumin

E. V. Chandrasekaran, Ram Chawda, Conrad Piskorz, Robert D. Locke, Alyssa Ta, Sharad A Ghamande, Kunle Odunsi, Shashikant Lele, Khushi L. Matta

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Affinity Gel-UDP was utilized to purify GlcNAc:β1,4Gal/GalNAc transferases (Ts) from human lymphoma spleen, ovarian tumor, and ovarian cancer sera. Mn2+ was found to be an absolute requirement for activity. Two molecular species containing both β1,4Gal/GalNAc-T activities were discernible when the purified ovarian tumor microsomal enzyme was subjected to Sephacryl S-100 HR column chromatography as well as native polyacylamide gel-electrophoresis. Acceptor specificity studies of the affinity-purified lymphoma spleen and ovarian tumor microsomal enzymes and the conventionally purified, as well as the cloned, bovine milk GlcNAc:β1,4Gal-Ts using a number of synthetic acceptors showed that the β(1,6)-linked GlcNAc moiety to α-GalNAc was the most efficient acceptor. As compared to the purified milk enzyme, the recombinant form exhibited sixfold GlcNAc:β1,4 GalNAc-T activity and up to eightfold GlcNAc6SO3β-:β1,4Gal-T activity. Further, the recombinant enzyme catalyzed the transfer of GalNAc to the terminal β-linked GlcNAc6SO3 moiety. Alpha-lactalbumin (α-LA) inhibited up to 85%, the transfer of Gal to the GlcNAc moiety linked either to Man or GlcNAc. On the contrary, α-LA had no significant influence on the transfer of GalNAc to the above acceptors. α-LA had no appreciable effect on the recombinant enzyme, except for the transfer of Gal or GalNAc to Glc. Both α- and β-glucosides, as well as α-N-acetylglucosaminide, did not serve as acceptors.

Original languageEnglish (US)
Pages (from-to)105-118
Number of pages14
JournalCarbohydrate Research
Volume334
Issue number2
DOIs
StatePublished - Aug 23 2001
Externally publishedYes

Fingerprint

Splenic Neoplasms
Enzootic Bovine Leukosis
Lactalbumin
Ovarian Neoplasms
Tumors
Milk
Enzymes
Neoplasms
Lymphoma
Spleen
Gels
Column chromatography
Uridine Diphosphate
Glucosides
Transferases
Electrophoresis
Chromatography
N-acetylglucopyranosylamine
polypeptide N-acetylgalactosaminyltransferase
Serum

Keywords

  • Bovine milk
  • Cancer sera
  • GlcNAc:β1,4Gal/GalNAc transferases
  • Human ovarian tumor
  • Kinetic properties
  • Lymphoma-spleen
  • Specificities

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Organic Chemistry

Cite this

Human ovarian cancer, lymphoma spleen, and bovine milk GlcNAc:β1,4Gal/GalNAc transferases : Two molecular species in ovarian tumor and induction of GalNAcβ1,4Glc synthesis by α-lactalbumin. / Chandrasekaran, E. V.; Chawda, Ram; Piskorz, Conrad; Locke, Robert D.; Ta, Alyssa; Ghamande, Sharad A; Odunsi, Kunle; Lele, Shashikant; Matta, Khushi L.

In: Carbohydrate Research, Vol. 334, No. 2, 23.08.2001, p. 105-118.

Research output: Contribution to journalArticle

Chandrasekaran, E. V. ; Chawda, Ram ; Piskorz, Conrad ; Locke, Robert D. ; Ta, Alyssa ; Ghamande, Sharad A ; Odunsi, Kunle ; Lele, Shashikant ; Matta, Khushi L. / Human ovarian cancer, lymphoma spleen, and bovine milk GlcNAc:β1,4Gal/GalNAc transferases : Two molecular species in ovarian tumor and induction of GalNAcβ1,4Glc synthesis by α-lactalbumin. In: Carbohydrate Research. 2001 ; Vol. 334, No. 2. pp. 105-118.
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abstract = "Affinity Gel-UDP was utilized to purify GlcNAc:β1,4Gal/GalNAc transferases (Ts) from human lymphoma spleen, ovarian tumor, and ovarian cancer sera. Mn2+ was found to be an absolute requirement for activity. Two molecular species containing both β1,4Gal/GalNAc-T activities were discernible when the purified ovarian tumor microsomal enzyme was subjected to Sephacryl S-100 HR column chromatography as well as native polyacylamide gel-electrophoresis. Acceptor specificity studies of the affinity-purified lymphoma spleen and ovarian tumor microsomal enzymes and the conventionally purified, as well as the cloned, bovine milk GlcNAc:β1,4Gal-Ts using a number of synthetic acceptors showed that the β(1,6)-linked GlcNAc moiety to α-GalNAc was the most efficient acceptor. As compared to the purified milk enzyme, the recombinant form exhibited sixfold GlcNAc:β1,4 GalNAc-T activity and up to eightfold GlcNAc6SO3β-:β1,4Gal-T activity. Further, the recombinant enzyme catalyzed the transfer of GalNAc to the terminal β-linked GlcNAc6SO3 moiety. Alpha-lactalbumin (α-LA) inhibited up to 85{\%}, the transfer of Gal to the GlcNAc moiety linked either to Man or GlcNAc. On the contrary, α-LA had no significant influence on the transfer of GalNAc to the above acceptors. α-LA had no appreciable effect on the recombinant enzyme, except for the transfer of Gal or GalNAc to Glc. Both α- and β-glucosides, as well as α-N-acetylglucosaminide, did not serve as acceptors.",
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AU - Chandrasekaran, E. V.

AU - Chawda, Ram

AU - Piskorz, Conrad

AU - Locke, Robert D.

AU - Ta, Alyssa

AU - Ghamande, Sharad A

AU - Odunsi, Kunle

AU - Lele, Shashikant

AU - Matta, Khushi L.

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AB - Affinity Gel-UDP was utilized to purify GlcNAc:β1,4Gal/GalNAc transferases (Ts) from human lymphoma spleen, ovarian tumor, and ovarian cancer sera. Mn2+ was found to be an absolute requirement for activity. Two molecular species containing both β1,4Gal/GalNAc-T activities were discernible when the purified ovarian tumor microsomal enzyme was subjected to Sephacryl S-100 HR column chromatography as well as native polyacylamide gel-electrophoresis. Acceptor specificity studies of the affinity-purified lymphoma spleen and ovarian tumor microsomal enzymes and the conventionally purified, as well as the cloned, bovine milk GlcNAc:β1,4Gal-Ts using a number of synthetic acceptors showed that the β(1,6)-linked GlcNAc moiety to α-GalNAc was the most efficient acceptor. As compared to the purified milk enzyme, the recombinant form exhibited sixfold GlcNAc:β1,4 GalNAc-T activity and up to eightfold GlcNAc6SO3β-:β1,4Gal-T activity. Further, the recombinant enzyme catalyzed the transfer of GalNAc to the terminal β-linked GlcNAc6SO3 moiety. Alpha-lactalbumin (α-LA) inhibited up to 85%, the transfer of Gal to the GlcNAc moiety linked either to Man or GlcNAc. On the contrary, α-LA had no significant influence on the transfer of GalNAc to the above acceptors. α-LA had no appreciable effect on the recombinant enzyme, except for the transfer of Gal or GalNAc to Glc. Both α- and β-glucosides, as well as α-N-acetylglucosaminide, did not serve as acceptors.

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