Chemistry in Living Cells: Detection of Active Proteasomes by a Two-Step Labeling Strategy†
Huib Ovaa Dr.
Department of Pathology, Harvard Medical School, 200 Longwood Avenue, Boston MA 02115, USA
These authors contributed equally.
Search for more papers by this authorPaul F. van Swieten
Gorlaeus Laboratories, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands, Fax: (+31) 71-527-4307
These authors contributed equally.
Search for more papers by this authorBenedikt M. Kessler Dr.
Department of Pathology, Harvard Medical School, 200 Longwood Avenue, Boston MA 02115, USA
Search for more papers by this authorMichiel A. Leeuwenburgh Dr.
Gorlaeus Laboratories, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands, Fax: (+31) 71-527-4307
Search for more papers by this authorEdda Fiebiger Dr.
Department of Pathology, Harvard Medical School, 200 Longwood Avenue, Boston MA 02115, USA
Search for more papers by this authorAdrianus M. C. H. van den Nieuwendijk
Gorlaeus Laboratories, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands, Fax: (+31) 71-527-4307
Search for more papers by this authorPaul J. Galardy Dr.
Department of Pathology, Harvard Medical School, 200 Longwood Avenue, Boston MA 02115, USA
Search for more papers by this authorGijsbert A. van der Marel Dr.
Gorlaeus Laboratories, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands, Fax: (+31) 71-527-4307
Search for more papers by this authorHidde L. Ploegh Prof. Dr.
Department of Pathology, Harvard Medical School, 200 Longwood Avenue, Boston MA 02115, USA
Search for more papers by this authorHerman S. Overkleeft Prof. Dr.
Gorlaeus Laboratories, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands, Fax: (+31) 71-527-4307
Search for more papers by this authorHuib Ovaa Dr.
Department of Pathology, Harvard Medical School, 200 Longwood Avenue, Boston MA 02115, USA
These authors contributed equally.
Search for more papers by this authorPaul F. van Swieten
Gorlaeus Laboratories, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands, Fax: (+31) 71-527-4307
These authors contributed equally.
Search for more papers by this authorBenedikt M. Kessler Dr.
Department of Pathology, Harvard Medical School, 200 Longwood Avenue, Boston MA 02115, USA
Search for more papers by this authorMichiel A. Leeuwenburgh Dr.
Gorlaeus Laboratories, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands, Fax: (+31) 71-527-4307
Search for more papers by this authorEdda Fiebiger Dr.
Department of Pathology, Harvard Medical School, 200 Longwood Avenue, Boston MA 02115, USA
Search for more papers by this authorAdrianus M. C. H. van den Nieuwendijk
Gorlaeus Laboratories, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands, Fax: (+31) 71-527-4307
Search for more papers by this authorPaul J. Galardy Dr.
Department of Pathology, Harvard Medical School, 200 Longwood Avenue, Boston MA 02115, USA
Search for more papers by this authorGijsbert A. van der Marel Dr.
Gorlaeus Laboratories, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands, Fax: (+31) 71-527-4307
Search for more papers by this authorHidde L. Ploegh Prof. Dr.
Department of Pathology, Harvard Medical School, 200 Longwood Avenue, Boston MA 02115, USA
Search for more papers by this authorHerman S. Overkleeft Prof. Dr.
Gorlaeus Laboratories, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands, Fax: (+31) 71-527-4307
Search for more papers by this authorThis work is supported by NIH Grant GM 62502 to H.L.P and NIH Training Grant T32 HL07574 to P.J.G. E.F. is supported by the APART program of the Austrian Academy of Sciences. H.O., P.F.S., and H.S.O. are financially supported by the Netherlands Organization for Scientific Research (NWO).
Graphical Abstract
In vivo targeting of the proteasome: Probe 1 is a cell-permeable irreversible inhibitor that alkylates the active-site residues of the proteasome in a Michael fashion. After cell lysis, a biotin moiety is introduced by Staudinger ligation to yield construct 2. This strategy allows activity profiling of the catalytic activities of the proteasome in vivo.
Supporting Information
Supporting information for this article is available on the WWW under http://www.wiley-vch.de/contents/jc_2002/2003/z51314_s.pdf or from the author.
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