The presence of outer arm fucose residues on the N -glycans of tissue inhibitor of metalloproteinases-1 reduces its activity

Han Ie Kim, Radka Saldova, Jun Hyoung Park, Young Hun Lee, David J. Harvey, Mark R. Wormald, Kieran Wynne, Giuliano Elia, Hwa Jung Kim, Pauline M. Rudd, Seung-Taek Lee

Research output: Contribution to journalArticle

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Abstract

Tissue inhibitor of metalloproteinases-1 (TIMP-1) inhibits matrix metalloproteinases (MMPs) by binding at a 1:1 stoichiometry. Here we have shown the involvement of N-glycosylation in the MMP inhibitory ability of TIMP-1. TIMP-1, purified from HEK 293 cells overexpressing TIMP-1 (293 TIMP-1), showed less binding and inhibitory abilities to MMPs than TIMP-1 purified from fibroblasts or SF9 insect cells infected with TIMP-1 baculovirus. Following deglycosylation of TIMP-1, all forms of TIMP-1 showed similar levels of MMP binding and inhibition, suggesting that glycosylation is involved in the regulation of these TIMP-1 activities. Analysis of the N-glycan structures showed that SF9 TIMP-1 has the simplest N-glycan structures, followed by fibroblast TIMP-1 and 293 TIMP-1, in order of increasing complexity in their N-glycan structures. Further analyses showed that cleavage of outer arm fucose residues from the N-glycans of 293 TIMP-1 or knockdown of both FUT4 and FUT7 (which encode for fucosyltransferases that add outer arm fucose residues to N-glycans) enhanced the MMP-binding and catalytic abilities of 293 TIMP-1, bringing them up to the levels of the other TIMP-1. These results demonstrate that the ability of TIMP-1 to inhibit MMPs is at least in part regulated by outer arm fucosylation of its N-glycans.

Original languageEnglish
Pages (from-to)3547-3560
Number of pages14
JournalJournal of Proteome Research
Volume12
Issue number8
DOIs
Publication statusPublished - 2013 Aug 2

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Tissue Inhibitor of Metalloproteinase-1
Fucose
Polysaccharides
Matrix Metalloproteinases
Glycosylation
Fibroblasts
Fucosyltransferases
Matrix Metalloproteinase Inhibitors
Baculoviridae
HEK293 Cells

All Science Journal Classification (ASJC) codes

  • Chemistry(all)
  • Biochemistry

Cite this

Kim, Han Ie ; Saldova, Radka ; Park, Jun Hyoung ; Lee, Young Hun ; Harvey, David J. ; Wormald, Mark R. ; Wynne, Kieran ; Elia, Giuliano ; Kim, Hwa Jung ; Rudd, Pauline M. ; Lee, Seung-Taek. / The presence of outer arm fucose residues on the N -glycans of tissue inhibitor of metalloproteinases-1 reduces its activity. In: Journal of Proteome Research. 2013 ; Vol. 12, No. 8. pp. 3547-3560.
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title = "The presence of outer arm fucose residues on the N -glycans of tissue inhibitor of metalloproteinases-1 reduces its activity",
abstract = "Tissue inhibitor of metalloproteinases-1 (TIMP-1) inhibits matrix metalloproteinases (MMPs) by binding at a 1:1 stoichiometry. Here we have shown the involvement of N-glycosylation in the MMP inhibitory ability of TIMP-1. TIMP-1, purified from HEK 293 cells overexpressing TIMP-1 (293 TIMP-1), showed less binding and inhibitory abilities to MMPs than TIMP-1 purified from fibroblasts or SF9 insect cells infected with TIMP-1 baculovirus. Following deglycosylation of TIMP-1, all forms of TIMP-1 showed similar levels of MMP binding and inhibition, suggesting that glycosylation is involved in the regulation of these TIMP-1 activities. Analysis of the N-glycan structures showed that SF9 TIMP-1 has the simplest N-glycan structures, followed by fibroblast TIMP-1 and 293 TIMP-1, in order of increasing complexity in their N-glycan structures. Further analyses showed that cleavage of outer arm fucose residues from the N-glycans of 293 TIMP-1 or knockdown of both FUT4 and FUT7 (which encode for fucosyltransferases that add outer arm fucose residues to N-glycans) enhanced the MMP-binding and catalytic abilities of 293 TIMP-1, bringing them up to the levels of the other TIMP-1. These results demonstrate that the ability of TIMP-1 to inhibit MMPs is at least in part regulated by outer arm fucosylation of its N-glycans.",
author = "Kim, {Han Ie} and Radka Saldova and Park, {Jun Hyoung} and Lee, {Young Hun} and Harvey, {David J.} and Wormald, {Mark R.} and Kieran Wynne and Giuliano Elia and Kim, {Hwa Jung} and Rudd, {Pauline M.} and Seung-Taek Lee",
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Kim, HI, Saldova, R, Park, JH, Lee, YH, Harvey, DJ, Wormald, MR, Wynne, K, Elia, G, Kim, HJ, Rudd, PM & Lee, S-T 2013, 'The presence of outer arm fucose residues on the N -glycans of tissue inhibitor of metalloproteinases-1 reduces its activity', Journal of Proteome Research, vol. 12, no. 8, pp. 3547-3560. https://doi.org/10.1021/pr400276r

The presence of outer arm fucose residues on the N -glycans of tissue inhibitor of metalloproteinases-1 reduces its activity. / Kim, Han Ie; Saldova, Radka; Park, Jun Hyoung; Lee, Young Hun; Harvey, David J.; Wormald, Mark R.; Wynne, Kieran; Elia, Giuliano; Kim, Hwa Jung; Rudd, Pauline M.; Lee, Seung-Taek.

In: Journal of Proteome Research, Vol. 12, No. 8, 02.08.2013, p. 3547-3560.

Research output: Contribution to journalArticle

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T1 - The presence of outer arm fucose residues on the N -glycans of tissue inhibitor of metalloproteinases-1 reduces its activity

AU - Kim, Han Ie

AU - Saldova, Radka

AU - Park, Jun Hyoung

AU - Lee, Young Hun

AU - Harvey, David J.

AU - Wormald, Mark R.

AU - Wynne, Kieran

AU - Elia, Giuliano

AU - Kim, Hwa Jung

AU - Rudd, Pauline M.

AU - Lee, Seung-Taek

PY - 2013/8/2

Y1 - 2013/8/2

N2 - Tissue inhibitor of metalloproteinases-1 (TIMP-1) inhibits matrix metalloproteinases (MMPs) by binding at a 1:1 stoichiometry. Here we have shown the involvement of N-glycosylation in the MMP inhibitory ability of TIMP-1. TIMP-1, purified from HEK 293 cells overexpressing TIMP-1 (293 TIMP-1), showed less binding and inhibitory abilities to MMPs than TIMP-1 purified from fibroblasts or SF9 insect cells infected with TIMP-1 baculovirus. Following deglycosylation of TIMP-1, all forms of TIMP-1 showed similar levels of MMP binding and inhibition, suggesting that glycosylation is involved in the regulation of these TIMP-1 activities. Analysis of the N-glycan structures showed that SF9 TIMP-1 has the simplest N-glycan structures, followed by fibroblast TIMP-1 and 293 TIMP-1, in order of increasing complexity in their N-glycan structures. Further analyses showed that cleavage of outer arm fucose residues from the N-glycans of 293 TIMP-1 or knockdown of both FUT4 and FUT7 (which encode for fucosyltransferases that add outer arm fucose residues to N-glycans) enhanced the MMP-binding and catalytic abilities of 293 TIMP-1, bringing them up to the levels of the other TIMP-1. These results demonstrate that the ability of TIMP-1 to inhibit MMPs is at least in part regulated by outer arm fucosylation of its N-glycans.

AB - Tissue inhibitor of metalloproteinases-1 (TIMP-1) inhibits matrix metalloproteinases (MMPs) by binding at a 1:1 stoichiometry. Here we have shown the involvement of N-glycosylation in the MMP inhibitory ability of TIMP-1. TIMP-1, purified from HEK 293 cells overexpressing TIMP-1 (293 TIMP-1), showed less binding and inhibitory abilities to MMPs than TIMP-1 purified from fibroblasts or SF9 insect cells infected with TIMP-1 baculovirus. Following deglycosylation of TIMP-1, all forms of TIMP-1 showed similar levels of MMP binding and inhibition, suggesting that glycosylation is involved in the regulation of these TIMP-1 activities. Analysis of the N-glycan structures showed that SF9 TIMP-1 has the simplest N-glycan structures, followed by fibroblast TIMP-1 and 293 TIMP-1, in order of increasing complexity in their N-glycan structures. Further analyses showed that cleavage of outer arm fucose residues from the N-glycans of 293 TIMP-1 or knockdown of both FUT4 and FUT7 (which encode for fucosyltransferases that add outer arm fucose residues to N-glycans) enhanced the MMP-binding and catalytic abilities of 293 TIMP-1, bringing them up to the levels of the other TIMP-1. These results demonstrate that the ability of TIMP-1 to inhibit MMPs is at least in part regulated by outer arm fucosylation of its N-glycans.

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