Functions of the N-glycans of rat leukemia inhibitory factor expressed in chinese hamster ovary cells

Ken Sasai, Jun Ichi Aikawa, Sakura Saburi, Hideaki Tojo, Satoshi Tanaka, Tomoya Ogawa, Kunio Shiota

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3 Citations (Scopus)

Abstract

Leukemia inhibitory factor (LIF) is a pluripotent growth factor which acts in various cell systems. LIF is a glycoprotein containing six putative N-glycosylation sites. We established Chinese hamster ovary (CHO) cell lines to evaluate the biological roles of the N-glycosylation in rat LIF (rLIF). The bioactivity of rLIF was evaluated in two different bioassay systems using F9 and DA-1a cells. Employing site directed mutagenesis, six N-glycosylation-deficient LIF mutants were generated by replacing each asparagine residue (N) (at positions N9, N34, N63, N73, N96, and N116) by glutamine (Q). The resultant mutants showed similar activity in the bioassay using F9 cells. However, N34Q was about 3 times more potent than the wild-type rLIF in the assay using DA-1a cells. These findings suggest that the presence of N-glycan at N34 suppresses cell proliferation. In contrast, N63Q was about 2.5 times less potent than the wild-type rLIF indicating the pivotal role of N63 glycosylation for rLIF bioactivity. Taken together, our data suggest that the N-glycans of LIF play different roles depending on the cell line and that glycosylation of each specific residue contributes differently to its bioactivity.

Original languageEnglish
Pages (from-to)999-1003
Number of pages5
JournalJournal of Biochemistry
Volume124
Issue number5
Publication statusPublished - 1998 Nov
Externally publishedYes

Keywords

  • DA-1a
  • F9
  • Leukemia inhibitory factor
  • N-glycosylation
  • Recombinant protein

ASJC Scopus subject areas

  • Biochemistry

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  • Cite this

    Sasai, K., Aikawa, J. I., Saburi, S., Tojo, H., Tanaka, S., Ogawa, T., & Shiota, K. (1998). Functions of the N-glycans of rat leukemia inhibitory factor expressed in chinese hamster ovary cells. Journal of Biochemistry, 124(5), 999-1003.