Ecto-ganglioside-sialidase activity of herpes simplex virus-transformed hamster embryo fibroblasts

Cara Lynne Schengrund, Abraham Rosenberg, Mary Ann Repman

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Cellular location of ganglioside-sialidase activity was determined in confluent hamster embryo fibroblasts transformed with herpes simplex virus type 2. Approximately equal specific activities of ganglioside-sialidase activity were found to be associated with the crude lysosomal and crude plasma membrane fractions isolated from whole cell homogenates. Whole transformed cells hydrolyzed exogenous ganglioside substrate, suggesting a partial location of the cellular sialidase on the outer surface of the plasma membrane of these cells. Intact cells were treated with the diazonium salt of sulfanilic acid, a nonpenetrating reagent inhibitory to ecto-enzymes (DePierre, J. W., and M. L. Kamovsky. 1974. J. Biol. Chem. 249: 7111-7120). Cytoplasmic lactate dehydrogenase activity was not inhibited by this treatment, and mitochondrial succinate dehydrogenase activity was inhibited only 10%, indicating that intracellular enzymes were not affected. 5′-Nucleotidase activity was diminished 90%, and sialidase very rapidly lost 40% of its exogenously directed activity. These results show that, in herpes simplex virus- transformed fibroblasts, ganglioside-sialidase is both a lysosomal and a plasma membrane enzyme. The plasma membrane sialidase is capable of acting on endogenous plasma membrane sialolipids and also functions in the cultured transformed cell as an ecto-enzyme which can attack exogenous substrates.

Original languageEnglish (US)
Pages (from-to)555-561
Number of pages7
JournalJournal of Cell Biology
Volume70
Issue number3
DOIs
StatePublished - Sep 1 1976

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Gangliosides
Neuraminidase
Simplexvirus
Cricetinae
Embryonic Structures
Fibroblasts
Cell Membrane
Enzymes
5'-Nucleotidase
Human Herpesvirus 2
Succinate Dehydrogenase
Plasma Cells
L-Lactate Dehydrogenase
Cultured Cells

All Science Journal Classification (ASJC) codes

  • Cell Biology

Cite this

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title = "Ecto-ganglioside-sialidase activity of herpes simplex virus-transformed hamster embryo fibroblasts",
abstract = "Cellular location of ganglioside-sialidase activity was determined in confluent hamster embryo fibroblasts transformed with herpes simplex virus type 2. Approximately equal specific activities of ganglioside-sialidase activity were found to be associated with the crude lysosomal and crude plasma membrane fractions isolated from whole cell homogenates. Whole transformed cells hydrolyzed exogenous ganglioside substrate, suggesting a partial location of the cellular sialidase on the outer surface of the plasma membrane of these cells. Intact cells were treated with the diazonium salt of sulfanilic acid, a nonpenetrating reagent inhibitory to ecto-enzymes (DePierre, J. W., and M. L. Kamovsky. 1974. J. Biol. Chem. 249: 7111-7120). Cytoplasmic lactate dehydrogenase activity was not inhibited by this treatment, and mitochondrial succinate dehydrogenase activity was inhibited only 10{\%}, indicating that intracellular enzymes were not affected. 5′-Nucleotidase activity was diminished 90{\%}, and sialidase very rapidly lost 40{\%} of its exogenously directed activity. These results show that, in herpes simplex virus- transformed fibroblasts, ganglioside-sialidase is both a lysosomal and a plasma membrane enzyme. The plasma membrane sialidase is capable of acting on endogenous plasma membrane sialolipids and also functions in the cultured transformed cell as an ecto-enzyme which can attack exogenous substrates.",
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Ecto-ganglioside-sialidase activity of herpes simplex virus-transformed hamster embryo fibroblasts. / Schengrund, Cara Lynne; Rosenberg, Abraham; Repman, Mary Ann.

In: Journal of Cell Biology, Vol. 70, No. 3, 01.09.1976, p. 555-561.

Research output: Contribution to journalArticle

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