Advanced

Furin is a chemokine-modifying enzyme : In vitro and in vivo processing of CXCL10 generates a C-terminally truncated chemokine retaining full activity

Hensbergen, Paul J.; Verzijl, Dennis; Balog, Crina I.A.; Dijkman, Remco; Van Der Schors, Roel C.; Van Der Raaij-Helmer, Elizabeth M.H.; Van Der Plas, Mariena J.A. LU ; Leurs, Rob; Deelder, André M. and Smit, Martine J., et al. (2004) In Journal of Biological Chemistry 279(14). p.13402-13411
Abstract

Chemokines comprise a class of structurally related proteins that are involved in many aspects of leukocyte migration under basal and inflammatory conditions. In addition to the large number of genes, limited processing of these proteins by a variety of enzymes enhances the complexity of the total spectrum of chemokine variants. We have recently shown that the native chemokine CXCL10 is processed at the C terminus, thereby shedding the last four amino acids. The present study was performed to elucidate the mechanism in vivo and in vitro and to study the biological activity of this novel isoform of CXCL10. Using a combination of protein purification and mass spectrometric techniques, we show that the production of C-terminally truncated... (More)

Chemokines comprise a class of structurally related proteins that are involved in many aspects of leukocyte migration under basal and inflammatory conditions. In addition to the large number of genes, limited processing of these proteins by a variety of enzymes enhances the complexity of the total spectrum of chemokine variants. We have recently shown that the native chemokine CXCL10 is processed at the C terminus, thereby shedding the last four amino acids. The present study was performed to elucidate the mechanism in vivo and in vitro and to study the biological activity of this novel isoform of CXCL10. Using a combination of protein purification and mass spectrometric techniques, we show that the production of C-terminally truncated CXCL10 by primary keratinocytes is inhibited in vivo by a specific inhibitor of pro-protein convertases (e.g. furin) but not by inhibition of matrix metalloproteinases. Moreover, CXCL10 is processed by furin in vitro, which is abrogated by a mutation in the furin recognition site. Using GTRγS binding, Ca2+ mobilization, and chemotaxis assays, we demonstrate that the C-terminally truncated CXCL10 variant is a potent ligand for CXCR3. Moreover, the inverse agonist activity on the virally encoded receptor ORF74 and the direct antibacterial activity of CXCL10 are fully retained. Hence, we have identified furin as a novel chemokine-modifying enzyme in vitro and most probably also in vivo, generating a C-terminally truncated CXCL10, which fully retains its (inverse) agonistic properties.

(Less)
Please use this url to cite or link to this publication:
author
, et al. (More)
(Less)
publishing date
type
Contribution to journal
publication status
published
in
Journal of Biological Chemistry
volume
279
issue
14
pages
10 pages
publisher
ASBMB
external identifiers
  • scopus:11144358288
ISSN
0021-9258
DOI
10.1074/jbc.M312814200
language
English
LU publication?
no
id
20245602-c064-4019-abf6-aa5a618e67e2
date added to LUP
2018-01-15 10:58:31
date last changed
2018-11-21 21:37:21
@article{20245602-c064-4019-abf6-aa5a618e67e2,
  abstract     = {<p>Chemokines comprise a class of structurally related proteins that are involved in many aspects of leukocyte migration under basal and inflammatory conditions. In addition to the large number of genes, limited processing of these proteins by a variety of enzymes enhances the complexity of the total spectrum of chemokine variants. We have recently shown that the native chemokine CXCL10 is processed at the C terminus, thereby shedding the last four amino acids. The present study was performed to elucidate the mechanism in vivo and in vitro and to study the biological activity of this novel isoform of CXCL10. Using a combination of protein purification and mass spectrometric techniques, we show that the production of C-terminally truncated CXCL10 by primary keratinocytes is inhibited in vivo by a specific inhibitor of pro-protein convertases (e.g. furin) but not by inhibition of matrix metalloproteinases. Moreover, CXCL10 is processed by furin in vitro, which is abrogated by a mutation in the furin recognition site. Using GTRγS binding, Ca<sup>2+</sup> mobilization, and chemotaxis assays, we demonstrate that the C-terminally truncated CXCL10 variant is a potent ligand for CXCR3. Moreover, the inverse agonist activity on the virally encoded receptor ORF74 and the direct antibacterial activity of CXCL10 are fully retained. Hence, we have identified furin as a novel chemokine-modifying enzyme in vitro and most probably also in vivo, generating a C-terminally truncated CXCL10, which fully retains its (inverse) agonistic properties.</p>},
  author       = {Hensbergen, Paul J. and Verzijl, Dennis and Balog, Crina I.A. and Dijkman, Remco and Van Der Schors, Roel C. and Van Der Raaij-Helmer, Elizabeth M.H. and Van Der Plas, Mariena J.A. and Leurs, Rob and Deelder, André M. and Smit, Martine J. and Tensen, Cornelis P.},
  issn         = {0021-9258},
  language     = {eng},
  month        = {04},
  number       = {14},
  pages        = {13402--13411},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {Furin is a chemokine-modifying enzyme : In vitro and in vivo processing of CXCL10 generates a C-terminally truncated chemokine retaining full activity},
  url          = {http://dx.doi.org/10.1074/jbc.M312814200},
  volume       = {279},
  year         = {2004},
}