Oligosaccharyltransferase activity is markedly increased during differentiation of a nonfusing myoblast cell line

Stephen R. Grant, Joseph K. Welply, Eric N. Olson, William J. Lennarz

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

We have studied several aspects of glycoprotein synthesis in myoblast differentiation by using a nonfusing myoblast cell line, BC3H1. Previous studies showed that transfer of proliferating undifferentiated BC3H1 cells to mitogen-depleted medium results in the cells' withdrawal from the cell cycle and induction of a variety of muscle-specific gene products [E. N. Olson, L. Glaser, J. P. Merlie, R. Sebane, and J. Lindstrom (1983) J. Biol. Chem. 258, 13946-13953]. Because cell surface glycoproteins have been implicated in myoblast differentiation, in the present study we measured the amount of oligosaccharyltransferase in microsomes isolated from BC3H1 cells at various stages of differentiation. By using an acceptor peptide containing the sequence AsnLeuThr, enzyme activity was measured by formation of [3H]glycopeptide. In addition, active enzyme protein was measured with a 125I-labeled photoreactive derivative of the acceptor tripeptide. Both of these independent assay methods revealed a marked increase in oligosaccharyltransferase when differentiation was induced by serum depletion. Moreover, mitogenic stimulation of differentiated cells resulted in a return of oligosaccharyltransferase to near basal levels. This reversible increase in this key enzyme in protein glycosylation occurred despite the fact that both total protein and glycoprotein synthesis were depressed during differentiation. These data indicate that during myogenesis the level of oligosaccharyltransferase is regulated in parallel with a number of muscle-specific gene products. These results are discussed in the context of regulation of the pathway of glycoprotein synthesis.

Original languageEnglish (US)
Pages (from-to)424-428
Number of pages5
JournalArchives of Biochemistry and Biophysics
Volume248
Issue number1
DOIs
StatePublished - 1986

Fingerprint

Myoblasts
Cells
Cell Line
Glycoproteins
Muscle
Enzymes
Genes
Glycosylation
Muscles
Proteins
Glycopeptides
Muscle Development
Membrane Glycoproteins
Enzyme activity
Microsomes
Mitogens
Assays
Cell Cycle
Derivatives
Peptides

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Oligosaccharyltransferase activity is markedly increased during differentiation of a nonfusing myoblast cell line. / Grant, Stephen R.; Welply, Joseph K.; Olson, Eric N.; Lennarz, William J.

In: Archives of Biochemistry and Biophysics, Vol. 248, No. 1, 1986, p. 424-428.

Research output: Contribution to journalArticle

@article{f151b406c4ea435aa35e4a0af4b96bdc,
title = "Oligosaccharyltransferase activity is markedly increased during differentiation of a nonfusing myoblast cell line",
abstract = "We have studied several aspects of glycoprotein synthesis in myoblast differentiation by using a nonfusing myoblast cell line, BC3H1. Previous studies showed that transfer of proliferating undifferentiated BC3H1 cells to mitogen-depleted medium results in the cells' withdrawal from the cell cycle and induction of a variety of muscle-specific gene products [E. N. Olson, L. Glaser, J. P. Merlie, R. Sebane, and J. Lindstrom (1983) J. Biol. Chem. 258, 13946-13953]. Because cell surface glycoproteins have been implicated in myoblast differentiation, in the present study we measured the amount of oligosaccharyltransferase in microsomes isolated from BC3H1 cells at various stages of differentiation. By using an acceptor peptide containing the sequence AsnLeuThr, enzyme activity was measured by formation of [3H]glycopeptide. In addition, active enzyme protein was measured with a 125I-labeled photoreactive derivative of the acceptor tripeptide. Both of these independent assay methods revealed a marked increase in oligosaccharyltransferase when differentiation was induced by serum depletion. Moreover, mitogenic stimulation of differentiated cells resulted in a return of oligosaccharyltransferase to near basal levels. This reversible increase in this key enzyme in protein glycosylation occurred despite the fact that both total protein and glycoprotein synthesis were depressed during differentiation. These data indicate that during myogenesis the level of oligosaccharyltransferase is regulated in parallel with a number of muscle-specific gene products. These results are discussed in the context of regulation of the pathway of glycoprotein synthesis.",
author = "Grant, {Stephen R.} and Welply, {Joseph K.} and Olson, {Eric N.} and Lennarz, {William J.}",
year = "1986",
doi = "10.1016/0003-9861(86)90439-X",
language = "English (US)",
volume = "248",
pages = "424--428",
journal = "Archives of Biochemistry and Biophysics",
issn = "0003-9861",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Oligosaccharyltransferase activity is markedly increased during differentiation of a nonfusing myoblast cell line

AU - Grant, Stephen R.

AU - Welply, Joseph K.

AU - Olson, Eric N.

AU - Lennarz, William J.

PY - 1986

Y1 - 1986

N2 - We have studied several aspects of glycoprotein synthesis in myoblast differentiation by using a nonfusing myoblast cell line, BC3H1. Previous studies showed that transfer of proliferating undifferentiated BC3H1 cells to mitogen-depleted medium results in the cells' withdrawal from the cell cycle and induction of a variety of muscle-specific gene products [E. N. Olson, L. Glaser, J. P. Merlie, R. Sebane, and J. Lindstrom (1983) J. Biol. Chem. 258, 13946-13953]. Because cell surface glycoproteins have been implicated in myoblast differentiation, in the present study we measured the amount of oligosaccharyltransferase in microsomes isolated from BC3H1 cells at various stages of differentiation. By using an acceptor peptide containing the sequence AsnLeuThr, enzyme activity was measured by formation of [3H]glycopeptide. In addition, active enzyme protein was measured with a 125I-labeled photoreactive derivative of the acceptor tripeptide. Both of these independent assay methods revealed a marked increase in oligosaccharyltransferase when differentiation was induced by serum depletion. Moreover, mitogenic stimulation of differentiated cells resulted in a return of oligosaccharyltransferase to near basal levels. This reversible increase in this key enzyme in protein glycosylation occurred despite the fact that both total protein and glycoprotein synthesis were depressed during differentiation. These data indicate that during myogenesis the level of oligosaccharyltransferase is regulated in parallel with a number of muscle-specific gene products. These results are discussed in the context of regulation of the pathway of glycoprotein synthesis.

AB - We have studied several aspects of glycoprotein synthesis in myoblast differentiation by using a nonfusing myoblast cell line, BC3H1. Previous studies showed that transfer of proliferating undifferentiated BC3H1 cells to mitogen-depleted medium results in the cells' withdrawal from the cell cycle and induction of a variety of muscle-specific gene products [E. N. Olson, L. Glaser, J. P. Merlie, R. Sebane, and J. Lindstrom (1983) J. Biol. Chem. 258, 13946-13953]. Because cell surface glycoproteins have been implicated in myoblast differentiation, in the present study we measured the amount of oligosaccharyltransferase in microsomes isolated from BC3H1 cells at various stages of differentiation. By using an acceptor peptide containing the sequence AsnLeuThr, enzyme activity was measured by formation of [3H]glycopeptide. In addition, active enzyme protein was measured with a 125I-labeled photoreactive derivative of the acceptor tripeptide. Both of these independent assay methods revealed a marked increase in oligosaccharyltransferase when differentiation was induced by serum depletion. Moreover, mitogenic stimulation of differentiated cells resulted in a return of oligosaccharyltransferase to near basal levels. This reversible increase in this key enzyme in protein glycosylation occurred despite the fact that both total protein and glycoprotein synthesis were depressed during differentiation. These data indicate that during myogenesis the level of oligosaccharyltransferase is regulated in parallel with a number of muscle-specific gene products. These results are discussed in the context of regulation of the pathway of glycoprotein synthesis.

UR - http://www.scopus.com/inward/record.url?scp=0022505121&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0022505121&partnerID=8YFLogxK

U2 - 10.1016/0003-9861(86)90439-X

DO - 10.1016/0003-9861(86)90439-X

M3 - Article

C2 - 3729427

AN - SCOPUS:0022505121

VL - 248

SP - 424

EP - 428

JO - Archives of Biochemistry and Biophysics

JF - Archives of Biochemistry and Biophysics

SN - 0003-9861

IS - 1

ER -