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 language | English |
|---|---|
| Pages (from-to) | 999-1003 |
| Number of pages | 5 |
| Journal | Journal of Biochemistry |
| Volume | 124 |
| Issue number | 5 |
| Publication status | Published - 1998 Nov |
| Externally published | Yes |
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Keywords
- DA-1a
- F9
- Leukemia inhibitory factor
- N-glycosylation
- Recombinant protein
ASJC Scopus subject areas
- Biochemistry
Cite this
Functions of the N-glycans of rat leukemia inhibitory factor expressed in chinese hamster ovary cells. / Sasai, Ken; Aikawa, Jun Ichi; Saburi, Sakura; Tojo, Hideaki; Tanaka, Satoshi; Ogawa, Tomoya; Shiota, Kunio.
In: Journal of Biochemistry, Vol. 124, No. 5, 11.1998, p. 999-1003.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Functions of the N-glycans of rat leukemia inhibitory factor expressed in chinese hamster ovary cells
AU - Sasai, Ken
AU - Aikawa, Jun Ichi
AU - Saburi, Sakura
AU - Tojo, Hideaki
AU - Tanaka, Satoshi
AU - Ogawa, Tomoya
AU - Shiota, Kunio
PY - 1998/11
Y1 - 1998/11
N2 - 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.
AB - 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.
KW - DA-1a
KW - F9
KW - Leukemia inhibitory factor
KW - N-glycosylation
KW - Recombinant protein
UR - http://www.scopus.com/inward/record.url?scp=0031787370&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0031787370&partnerID=8YFLogxK
M3 - Article
VL - 124
SP - 999
EP - 1003
JO - Journal of Biochemistry
JF - Journal of Biochemistry
SN - 0021-924X
IS - 5
ER -