Signaling Pathways Underlying Eosinophil Cell Motility Revealed by Using Caged Peptides
Insights into structure-function relations of many proteins opens the possibility of engineering peptides to selectively interfere with a protein's activity. To facilitate the use of peptides as probes of cellular processes, we have developed caged peptides whose influence on specific proteins...
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Published in | Proceedings of the National Academy of Sciences - PNAS Vol. 95; no. 4; pp. 1568 - 1573 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
National Academy of Sciences of the United States of America
17.02.1998
National Acad Sciences National Academy of Sciences The National Academy of Sciences |
Subjects | |
Online Access | Get full text |
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Summary: | Insights into structure-function relations of many proteins opens the possibility of engineering peptides to selectively interfere with a protein's activity. To facilitate the use of peptides as probes of cellular processes, we have developed caged peptides whose influence on specific proteins can be suddenly and uniformly changed by near-UV light. Two peptides are described which, on photolysis of a caging moiety, block the action of calcium-calmodulin or myosin light chain kinase (MLCK). The efficacy of theses peptides is demonstrated in vitro and in vivo by determining their effect before and after photolysis on activities of isolated enzymes and cellular functions known to depend on calcium-calmodulin and MLCK. These caged peptides each were injected into motile, polarized eosinophils, and when exposed to light promptly blocked cell locomotion in a similar manner. The results indicate that the action of calcium-calmodulin and MLCK, and by inference myosin II, are required for the ameboid locomotion of these cells. This methodology provides a powerful means for assessing the role of these and other proteins in a wide range of spatio-temporally complex functions in intact living cells. |
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Bibliography: | ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2 Edited by Thomas D. Pollard, Salk Institute for Biological Studies, La Jolla, CA, and approved December 22, 1997 To whom reprint requests should be addressed at: Department of Physiology, University of Wisconsin School of Medicine, 1300 University Avenue, Madison, WI 53706. e-mail: jwwalker@facstaff.wisc.edu. |
ISSN: | 0027-8424 1091-6490 |
DOI: | 10.1073/pnas.95.4.1568 |