Increased expression of vascular cell adhesion molecule 1 (VCAM1) is associated with a variety of chronic inflammatory conditions, making its expression and function a target for therapeutic intervention1,2,3. We have recently identified CAM741, a derivative of a fungus-derived cyclopeptolide that acts as a selective inhibitor of VCAM1 synthesis in endothelial cells. Here we show that the compound represses the biosynthesis of VCAM1 in cells by blocking the process of cotranslational translocation, which is dependent on the signal peptide of VCAM1. CAM741 does not inhibit targeting of the VCAM1 nascent chains to the translocon channel but prevents translocation to the luminal side of the endoplasmic reticulum (ER), through a process that involves the translocon component Sec61β. Consequently, the VCAM1 precursor protein is synthesized towards the cytosolic compartment of the cells, where it is degraded. Our results indicate that the inhibition of cotranslational translocation with low-molecular-mass compounds, using specificity conferred by signal peptides, can modulate the biosynthesis of certain secreted and/or membrane proteins. In addition, they highlight cotranslational translocation at the ER membrane as a potential target for drug discovery.
Subscribe to Journal
Get full journal access for 1 year
only $3.90 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Springer, T. A. Traffic signals on endothelium for lymphocyte recirculation and leukocyte emigration. Annu. Rev. Physiol. 57, 827–872 (1995)
Schreiner, E. P., Oberhauser, B. & Foster, C. A. Inhibitors of vascular cell adhesion molecule-1 expression. Expert Opin. Therapeut. Patents 13, 149–166 (2003)
Bevilacqua, M. P., Nelson, R. M., Mannori, G. & Cecconi, O. Endothelial-leukocyte adhesion molecules in human disease. Annu. Rev. Med. 45, 361–378 (1994)
Boger, D. L., Keim, H., Oberhauser, B., Schreiner, E. P. & Foster, C. A. Total synthesis of HUN-7293. J. Am. Chem. Soc. 121, 6197–6205 (1999)
Hommel, U. et al. The 3D-structure of a natural inhibitor of cell adhesion molecule expression. FEBS Lett. 379, 69–73 (1996)
Walter, P. & Johnson, A. E. Signal sequence recognition and protein targeting to the endoplasmic reticulum membrane. Annu. Rev. Cell Biol. 10, 87–119 (1994)
Matlack, K. E., Mothes, W. & Rapoport, T. A. Protein translocation: tunnel vision. Cell 92, 381–390 (1998)
Johnson, A. E. & van Waes, M. A. The translocon: a dynamic gateway at the ER membrane. Annu. Rev. Cell Dev. Biol. 15, 799–842 (1999)
Jungnickel, B. & Rapoport, T. A. A posttargeting signal sequence recognition event in the endoplasmic reticulum membrane. Cell 82, 261–270 (1995)
Kim, S. J., Mitra, D., Salerno, J. R. & Hegde, R. S. Signal sequences control gating of the protein translocation channel in a substrate-specific manner. Dev. Cell 2, 207–217 (2002)
Rutkowski, D. T., Lingappa, V. R. & Hegde, R. S. Substrate-specific regulation of the ribosome–translocon junction by N-terminal signal sequences. Proc. Natl Acad. Sci. USA 98, 7823–7828 (2001)
Fons, R. D., Bogert, B. A. & Hegde, R. S. Substrate-specific function of the translocon-associated protein complex during translocation across the ER membrane. J. Cell Biol. 160, 529–539 (2003)
Mothes, W., Prehn, S. & Rapoport, T. A. Systematic probing of the environment of a translocating secretory protein during translocation through the ER membrane. EMBO J. 13, 3973–3982 (1994)
Kalies, K. U., Rapoport, T. A. & Hartmann, E. The beta subunit of the Sec61 complex facilitates cotranslational protein transport and interacts with the signal peptidase during translocation. J. Cell Biol. 141, 887–894 (1998)
DeMartino, G. N. & Slaughter, C. A. The proteasome, a novel protease regulated by multiple mechanisms. J. Biol. Chem. 274, 22123–22126 (1999)
We thank A. Rot for generation of the Sec61β antiserum, H. Jaksche for its purification, and B. Kappel, H. Pertl, L. Hofer and N. Lettner for technical assistance.
H.H., S.W., C.A.F., E.P.S., B.O., J.E.d.V. and I.J.D.L. are employed by Novartis. In addition, B.O., J.E.d.V. and I.J.D.L. own restricted stocks. J.B., K.M. and C.D.-N. left Novartis and do not have any conflicting financial interests.
Data demonstrating that the VCAM-1 SP plus the first amino acid of the VCAM-1 mature region are required for full sensitivity to CAM741, while parts of the N-terminal region of the VCAM-1 SP are dispensable for translocation and sensitivity. (DOC 24 kb)
In vitro translocation experiments described in Supplementary Data. (PDF 225 kb)
Results from transient transfection experiments described in Supplementary Data. (DOC 32 kb)
Methods for experiment shown in Supplementary Table S1. (DOC 19 kb)
About this article
Cite this article
Besemer, J., Harant, H., Wang, S. et al. Selective inhibition of cotranslational translocation of vascular cell adhesion molecule 1. Nature 436, 290–293 (2005) doi:10.1038/nature03670
Vascular Pharmacology (2019)
Oxidative Stress Induced by the Deubiquitinase Inhibitor b-AP15 Is Associated with Mitochondrial Impairment
Oxidative Medicine and Cellular Longevity (2019)
Current Pharmaceutical Design (2019)
Preprotein signature for full susceptibility to the co‐translational translocation inhibitor cyclotriazadisulfonamide
FEBS Letters (2019)