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Published 2006 | Supplemental Material
Journal Article Open

Fluorescence Visualization of Newly Synthesized Proteins in Mammalian Cells

Abstract

Modern proteomic methods enable efficient identification of the hundreds or thousands of proteins present in whole cells or in isolated organelles. However, a thorough understanding of the proteome requires insight into protein localization as well as protein identity. Recently, visualization of newly synthesized proteins in bacterial cells was demonstrated through co-translational introduction of an alkynyl amino acid followed by selective CuI-catalyzed ligation of the alkynyl side chain to the fluorogenic dye 3-azido-7-hydroxycoumarin. Here we report that selective fluorescence labeling and imaging of newly synthesized proteins can be accomplished in a diverse set of mammalian cells.

Additional Information

© 2006 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Received: May 26, 2006 Revised: August 1, 2006 Published online: October 11, 2006 We thank Scott Fraser, Chris Waters, and the Beckman Imaging Center for advice on microscopy, and Rochelle Diamond, Stephanie Adams, and the Caltech Flow Cytometry Facility for assistance with flow cytometry. We thank Anand Asthagiri, Chase Beisel, David Chan, Scott Detmer, Nicholas Graham, Melissa Pope, and Christina Smolke for cell lines and reagents. James Van Deventer made helpful comments on the manuscript. This work was supported by a Fannie and John Hertz Foundation Fellowship (to K.E.B.), a Whitaker Foundation Graduate Fellowship (to J.C.L.), a postdoctoral fellowship from the German Academy for Natural Scientists Leopoldina (BMBF-LPD9901/8-95 to D.C.D.), an NSF-NER grant (to Q.W.), NIH grant GM62523 (to D.A.T.), and the Beckman Institute at Caltech. E.M.S. is an Investigator of the Howard Hughes Medical Institute.

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Created:
August 19, 2023
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October 19, 2023