Furin proteolytically processes the heparin-binding region of extracellular superoxide dismutase.

Russell P Bowler, Mike Nicks, Dorte Aa Olsen, Ida B Thøgersen, Zuzana Valnickova, Peter Højrup, Alex Franzusoff, Jan J Enghild, James D Crapo

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Extracellular superoxide dismutase (EC-SOD) is an antioxidant enzyme that attenuates brain and lung injury from oxidative stress. A polybasic region in the carboxyl terminus distinguishes EC-SOD from other superoxide dismutases and determines EC-SOD's tissue half-life and affinity for heparin. There are two types of EC-SOD that differ based on the presence or absence of this heparin-binding region. It has recently been shown that proteolytic removal of the heparin-binding region is an intracellular event (Enghild, J. J., Thogersen, I. B., Oury, T. D., Valnickova, Z., Hojrup, P., and Crapo, J. D. (1999) J. Biol. Chem. 274, 14818-14822). By using mammalian cell lines, we have now determined that removal of the heparin-binding region occurs after passage through the Golgi network but before being secreted into the extracellular space. Specific protease inhibitors and overexpression of intracellular proteases implicate furin as a processing protease. In vitro experiments using furin and purified EC-SOD suggest that furin proteolytically cleaves EC-SOD in the middle of the polybasic region and then requires an additional carboxypeptidase to remove the remaining lysines and arginines. A mutation in Arg(213) renders EC-SOD resistant to furin processing. These results indicate that furin-dependent processing of EC-SOD is important for determining the tissue distribution and half-life of EC-SOD.
Original languageEnglish
JournalJournal of Biological Chemistry
Volume277
Issue number19
Pages (from-to)16505-11
Number of pages6
ISSN0021-9258
DOIs
Publication statusPublished - 10. May 2002

Fingerprint

Furin
Superoxide Dismutase
Heparin
Half-Life
Peptide Hydrolases
Processing
Tissue
Carboxypeptidases
Oxidative stress
Protease Inhibitors
Lysine
Arginine
Brain
Antioxidants
Cells
Cell Line

Keywords

  • Amino Acid Sequence
  • Animals
  • Arginine
  • Blotting, Western
  • Brefeldin A
  • CHO Cells
  • Cell Line
  • Cricetinae
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Inhibitors
  • Furin
  • Glycine
  • Golgi Apparatus
  • Heparin
  • Humans
  • Mass Spectrometry
  • Mice
  • Molecular Sequence Data
  • Mutation
  • Oxidative Stress
  • Precipitin Tests
  • Protein Binding
  • Protein Structure, Tertiary
  • Protein Synthesis Inhibitors
  • Rats
  • Subtilisins
  • Superoxide Dismutase
  • Temperature
  • Time Factors
  • Tissue Distribution
  • Tumor Cells, Cultured

Cite this

Bowler, R. P., Nicks, M., Olsen, D. A., Thøgersen, I. B., Valnickova, Z., Højrup, P., ... Crapo, J. D. (2002). Furin proteolytically processes the heparin-binding region of extracellular superoxide dismutase. Journal of Biological Chemistry, 277(19), 16505-11. https://doi.org/10.1074/jbc.M105409200
Bowler, Russell P ; Nicks, Mike ; Olsen, Dorte Aa ; Thøgersen, Ida B ; Valnickova, Zuzana ; Højrup, Peter ; Franzusoff, Alex ; Enghild, Jan J ; Crapo, James D. / Furin proteolytically processes the heparin-binding region of extracellular superoxide dismutase. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 19. pp. 16505-11.
@article{0d8e11f062da11ddb1a1000ea68e967b,
title = "Furin proteolytically processes the heparin-binding region of extracellular superoxide dismutase.",
abstract = "Extracellular superoxide dismutase (EC-SOD) is an antioxidant enzyme that attenuates brain and lung injury from oxidative stress. A polybasic region in the carboxyl terminus distinguishes EC-SOD from other superoxide dismutases and determines EC-SOD's tissue half-life and affinity for heparin. There are two types of EC-SOD that differ based on the presence or absence of this heparin-binding region. It has recently been shown that proteolytic removal of the heparin-binding region is an intracellular event (Enghild, J. J., Thogersen, I. B., Oury, T. D., Valnickova, Z., Hojrup, P., and Crapo, J. D. (1999) J. Biol. Chem. 274, 14818-14822). By using mammalian cell lines, we have now determined that removal of the heparin-binding region occurs after passage through the Golgi network but before being secreted into the extracellular space. Specific protease inhibitors and overexpression of intracellular proteases implicate furin as a processing protease. In vitro experiments using furin and purified EC-SOD suggest that furin proteolytically cleaves EC-SOD in the middle of the polybasic region and then requires an additional carboxypeptidase to remove the remaining lysines and arginines. A mutation in Arg(213) renders EC-SOD resistant to furin processing. These results indicate that furin-dependent processing of EC-SOD is important for determining the tissue distribution and half-life of EC-SOD.",
keywords = "Amino Acid Sequence, Animals, Arginine, Blotting, Western, Brefeldin A, CHO Cells, Cell Line, Cricetinae, Electrophoresis, Polyacrylamide Gel, Enzyme Inhibitors, Furin, Glycine, Golgi Apparatus, Heparin, Humans, Mass Spectrometry, Mice, Molecular Sequence Data, Mutation, Oxidative Stress, Precipitin Tests, Protein Binding, Protein Structure, Tertiary, Protein Synthesis Inhibitors, Rats, Subtilisins, Superoxide Dismutase, Temperature, Time Factors, Tissue Distribution, Tumor Cells, Cultured",
author = "Bowler, {Russell P} and Mike Nicks and Olsen, {Dorte Aa} and Th{\o}gersen, {Ida B} and Zuzana Valnickova and Peter H{\o}jrup and Alex Franzusoff and Enghild, {Jan J} and Crapo, {James D}",
year = "2002",
month = "5",
day = "10",
doi = "10.1074/jbc.M105409200",
language = "English",
volume = "277",
pages = "16505--11",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology, Inc.",
number = "19",

}

Bowler, RP, Nicks, M, Olsen, DA, Thøgersen, IB, Valnickova, Z, Højrup, P, Franzusoff, A, Enghild, JJ & Crapo, JD 2002, 'Furin proteolytically processes the heparin-binding region of extracellular superoxide dismutase.', Journal of Biological Chemistry, vol. 277, no. 19, pp. 16505-11. https://doi.org/10.1074/jbc.M105409200

Furin proteolytically processes the heparin-binding region of extracellular superoxide dismutase. / Bowler, Russell P; Nicks, Mike; Olsen, Dorte Aa; Thøgersen, Ida B; Valnickova, Zuzana; Højrup, Peter; Franzusoff, Alex; Enghild, Jan J; Crapo, James D.

In: Journal of Biological Chemistry, Vol. 277, No. 19, 10.05.2002, p. 16505-11.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Furin proteolytically processes the heparin-binding region of extracellular superoxide dismutase.

AU - Bowler, Russell P

AU - Nicks, Mike

AU - Olsen, Dorte Aa

AU - Thøgersen, Ida B

AU - Valnickova, Zuzana

AU - Højrup, Peter

AU - Franzusoff, Alex

AU - Enghild, Jan J

AU - Crapo, James D

PY - 2002/5/10

Y1 - 2002/5/10

N2 - Extracellular superoxide dismutase (EC-SOD) is an antioxidant enzyme that attenuates brain and lung injury from oxidative stress. A polybasic region in the carboxyl terminus distinguishes EC-SOD from other superoxide dismutases and determines EC-SOD's tissue half-life and affinity for heparin. There are two types of EC-SOD that differ based on the presence or absence of this heparin-binding region. It has recently been shown that proteolytic removal of the heparin-binding region is an intracellular event (Enghild, J. J., Thogersen, I. B., Oury, T. D., Valnickova, Z., Hojrup, P., and Crapo, J. D. (1999) J. Biol. Chem. 274, 14818-14822). By using mammalian cell lines, we have now determined that removal of the heparin-binding region occurs after passage through the Golgi network but before being secreted into the extracellular space. Specific protease inhibitors and overexpression of intracellular proteases implicate furin as a processing protease. In vitro experiments using furin and purified EC-SOD suggest that furin proteolytically cleaves EC-SOD in the middle of the polybasic region and then requires an additional carboxypeptidase to remove the remaining lysines and arginines. A mutation in Arg(213) renders EC-SOD resistant to furin processing. These results indicate that furin-dependent processing of EC-SOD is important for determining the tissue distribution and half-life of EC-SOD.

AB - Extracellular superoxide dismutase (EC-SOD) is an antioxidant enzyme that attenuates brain and lung injury from oxidative stress. A polybasic region in the carboxyl terminus distinguishes EC-SOD from other superoxide dismutases and determines EC-SOD's tissue half-life and affinity for heparin. There are two types of EC-SOD that differ based on the presence or absence of this heparin-binding region. It has recently been shown that proteolytic removal of the heparin-binding region is an intracellular event (Enghild, J. J., Thogersen, I. B., Oury, T. D., Valnickova, Z., Hojrup, P., and Crapo, J. D. (1999) J. Biol. Chem. 274, 14818-14822). By using mammalian cell lines, we have now determined that removal of the heparin-binding region occurs after passage through the Golgi network but before being secreted into the extracellular space. Specific protease inhibitors and overexpression of intracellular proteases implicate furin as a processing protease. In vitro experiments using furin and purified EC-SOD suggest that furin proteolytically cleaves EC-SOD in the middle of the polybasic region and then requires an additional carboxypeptidase to remove the remaining lysines and arginines. A mutation in Arg(213) renders EC-SOD resistant to furin processing. These results indicate that furin-dependent processing of EC-SOD is important for determining the tissue distribution and half-life of EC-SOD.

KW - Amino Acid Sequence

KW - Animals

KW - Arginine

KW - Blotting, Western

KW - Brefeldin A

KW - CHO Cells

KW - Cell Line

KW - Cricetinae

KW - Electrophoresis, Polyacrylamide Gel

KW - Enzyme Inhibitors

KW - Furin

KW - Glycine

KW - Golgi Apparatus

KW - Heparin

KW - Humans

KW - Mass Spectrometry

KW - Mice

KW - Molecular Sequence Data

KW - Mutation

KW - Oxidative Stress

KW - Precipitin Tests

KW - Protein Binding

KW - Protein Structure, Tertiary

KW - Protein Synthesis Inhibitors

KW - Rats

KW - Subtilisins

KW - Superoxide Dismutase

KW - Temperature

KW - Time Factors

KW - Tissue Distribution

KW - Tumor Cells, Cultured

U2 - 10.1074/jbc.M105409200

DO - 10.1074/jbc.M105409200

M3 - Journal article

VL - 277

SP - 16505

EP - 16511

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 19

ER -