Activity probe for in vivo profiling of the specificity of proteasome inhibitor bortezomib

Celia R. Berkers, Martijn Verdoes, Eben Lichtman, Edda Fiebiger, Benedikt M. Kessler, Kenneth C. Anderson, Hidde L. Ploegh, Huib Ovaa, Paul J. Galardy

Research output: Contribution to journalArticle

184 Citations (Scopus)

Abstract

Proteasome inhibitors, such as the dipeptide boronic acid bortezomib, are emerging as important tools in the treatment of the fatal hematologic malignancy multiple myeloma. Despite the recent US Food and Drug Administration approval of bortezomib (PS341, Velcade) for the treatment of refractory multiple myeloma, many of the basic pharmacologic parameters of bortezomib and its mode of action on myeloma cells remain to be determined. We describe the synthesis and use of a cell-permeant active site-directed probe, which allows profiling of proteasomal activities in living cells. When we compared proteasome activity patterns in cultured cells and crude cell extracts with this probe, we observed substantial differences, stressing the importance for bioassays compatible with live cells to ensure accuracy of such measurements. Using this probe, we investigated the in vivo subunit specificities of bortezomib and another inhibitor, MG132.

Original languageEnglish (US)
Pages (from-to)357-362
Number of pages6
JournalNature Methods
Volume2
Issue number5
DOIs
StatePublished - May 2005
Externally publishedYes

Fingerprint

Proteasome Inhibitors
Multiple Myeloma
Cells
Boronic Acids
Drug Approval
Bioassay
Dipeptides
Hematologic Neoplasms
Proteasome Endopeptidase Complex
Cell Extracts
Complex Mixtures
Biological Assay
Refractory materials
Cultured Cells
Catalytic Domain
Bortezomib
Food

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology
  • Cell Biology

Cite this

Berkers, C. R., Verdoes, M., Lichtman, E., Fiebiger, E., Kessler, B. M., Anderson, K. C., ... Galardy, P. J. (2005). Activity probe for in vivo profiling of the specificity of proteasome inhibitor bortezomib. Nature Methods, 2(5), 357-362. https://doi.org/10.1038/nmeth759

Activity probe for in vivo profiling of the specificity of proteasome inhibitor bortezomib. / Berkers, Celia R.; Verdoes, Martijn; Lichtman, Eben; Fiebiger, Edda; Kessler, Benedikt M.; Anderson, Kenneth C.; Ploegh, Hidde L.; Ovaa, Huib; Galardy, Paul J.

In: Nature Methods, Vol. 2, No. 5, 05.2005, p. 357-362.

Research output: Contribution to journalArticle

Berkers, CR, Verdoes, M, Lichtman, E, Fiebiger, E, Kessler, BM, Anderson, KC, Ploegh, HL, Ovaa, H & Galardy, PJ 2005, 'Activity probe for in vivo profiling of the specificity of proteasome inhibitor bortezomib', Nature Methods, vol. 2, no. 5, pp. 357-362. https://doi.org/10.1038/nmeth759
Berkers CR, Verdoes M, Lichtman E, Fiebiger E, Kessler BM, Anderson KC et al. Activity probe for in vivo profiling of the specificity of proteasome inhibitor bortezomib. Nature Methods. 2005 May;2(5):357-362. https://doi.org/10.1038/nmeth759
Berkers, Celia R. ; Verdoes, Martijn ; Lichtman, Eben ; Fiebiger, Edda ; Kessler, Benedikt M. ; Anderson, Kenneth C. ; Ploegh, Hidde L. ; Ovaa, Huib ; Galardy, Paul J. / Activity probe for in vivo profiling of the specificity of proteasome inhibitor bortezomib. In: Nature Methods. 2005 ; Vol. 2, No. 5. pp. 357-362.
@article{d0ad36af748e4500b1a10572bc387ea4,
title = "Activity probe for in vivo profiling of the specificity of proteasome inhibitor bortezomib",
abstract = "Proteasome inhibitors, such as the dipeptide boronic acid bortezomib, are emerging as important tools in the treatment of the fatal hematologic malignancy multiple myeloma. Despite the recent US Food and Drug Administration approval of bortezomib (PS341, Velcade) for the treatment of refractory multiple myeloma, many of the basic pharmacologic parameters of bortezomib and its mode of action on myeloma cells remain to be determined. We describe the synthesis and use of a cell-permeant active site-directed probe, which allows profiling of proteasomal activities in living cells. When we compared proteasome activity patterns in cultured cells and crude cell extracts with this probe, we observed substantial differences, stressing the importance for bioassays compatible with live cells to ensure accuracy of such measurements. Using this probe, we investigated the in vivo subunit specificities of bortezomib and another inhibitor, MG132.",
author = "Berkers, {Celia R.} and Martijn Verdoes and Eben Lichtman and Edda Fiebiger and Kessler, {Benedikt M.} and Anderson, {Kenneth C.} and Ploegh, {Hidde L.} and Huib Ovaa and Galardy, {Paul J.}",
year = "2005",
month = "5",
doi = "10.1038/nmeth759",
language = "English (US)",
volume = "2",
pages = "357--362",
journal = "PLoS Medicine",
issn = "1549-1277",
publisher = "Nature Publishing Group",
number = "5",

}

TY - JOUR

T1 - Activity probe for in vivo profiling of the specificity of proteasome inhibitor bortezomib

AU - Berkers, Celia R.

AU - Verdoes, Martijn

AU - Lichtman, Eben

AU - Fiebiger, Edda

AU - Kessler, Benedikt M.

AU - Anderson, Kenneth C.

AU - Ploegh, Hidde L.

AU - Ovaa, Huib

AU - Galardy, Paul J.

PY - 2005/5

Y1 - 2005/5

N2 - Proteasome inhibitors, such as the dipeptide boronic acid bortezomib, are emerging as important tools in the treatment of the fatal hematologic malignancy multiple myeloma. Despite the recent US Food and Drug Administration approval of bortezomib (PS341, Velcade) for the treatment of refractory multiple myeloma, many of the basic pharmacologic parameters of bortezomib and its mode of action on myeloma cells remain to be determined. We describe the synthesis and use of a cell-permeant active site-directed probe, which allows profiling of proteasomal activities in living cells. When we compared proteasome activity patterns in cultured cells and crude cell extracts with this probe, we observed substantial differences, stressing the importance for bioassays compatible with live cells to ensure accuracy of such measurements. Using this probe, we investigated the in vivo subunit specificities of bortezomib and another inhibitor, MG132.

AB - Proteasome inhibitors, such as the dipeptide boronic acid bortezomib, are emerging as important tools in the treatment of the fatal hematologic malignancy multiple myeloma. Despite the recent US Food and Drug Administration approval of bortezomib (PS341, Velcade) for the treatment of refractory multiple myeloma, many of the basic pharmacologic parameters of bortezomib and its mode of action on myeloma cells remain to be determined. We describe the synthesis and use of a cell-permeant active site-directed probe, which allows profiling of proteasomal activities in living cells. When we compared proteasome activity patterns in cultured cells and crude cell extracts with this probe, we observed substantial differences, stressing the importance for bioassays compatible with live cells to ensure accuracy of such measurements. Using this probe, we investigated the in vivo subunit specificities of bortezomib and another inhibitor, MG132.

UR - http://www.scopus.com/inward/record.url?scp=18244406798&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=18244406798&partnerID=8YFLogxK

U2 - 10.1038/nmeth759

DO - 10.1038/nmeth759

M3 - Article

C2 - 15846363

AN - SCOPUS:18244406798

VL - 2

SP - 357

EP - 362

JO - PLoS Medicine

JF - PLoS Medicine

SN - 1549-1277

IS - 5

ER -