Heilshorn, Sarah C. and Liu, Julie C. and Tirrell, David A. (2005) Cell-Binding Domain Context Affects Cell Behavior on Engineered Proteins. Biomacromolecules, 6 (1). pp. 318-323. ISSN 1525-7797. doi:10.1021/bm049627q. https://resolver.caltech.edu/CaltechAUTHORS:20150114-121449592
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Abstract
A family of artificial extracellular matrix proteins developed for application in small-diameter vascular grafts is used to examine the importance of cell-binding domain context on cell adhesion and spreading. The engineered protein sequences are derived from the naturally occurring extracellular matrix proteins elastin and fibronectin. While each engineered protein contains identical CS5 cell-binding domain sequences, the lysine residues that serve as cross-linking sites are either (i) within the elastin cassettes or (ii) confined to the ends of the protein. Endothelial cells adhere specifically to the CS5 sequence in both of these proteins, but cell adhesion and spreading are more robust on proteins in which the lysine residues are confined to the terminal regions of the chain. These results may be due to altered protein conformations that affect either the accessibility of the CS5 sequence or its affinity for the α_4β_1 integrin receptor on the endothelial cell surface. Amino acid choice outside the cell-binding domain can thus have a significant impact on the behavior of cells cultured on artificial extracellular matrix proteins.
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Additional Information: | Copyright © 2005 American Chemical Society. Published In Issue January 10, 2005. Publication Date (Web): December 4, 2004. Received June 29, 2004. Revised Manuscript Received October 5, 2004. Acknowledgment. This work was supported by NIH Grant 5 R01 HL59987-03, NSF Grant BES-9901648, and a Whitaker graduate fellowship. We thank Kathleen Di Zio for helpful discussion regarding protein purification, Paul Nowatzki for providing protein samples, the Electron Microscopy Laboratory in the Biology Division at Caltech and Robert Strittmatter for help with the scanning electron micrographs, and Krystle Wang, Gustavo Olm, and Regina Wilpiszeski for help with the BCA and cell detachment experiments. Supporting Information Available. Supporting figures include 1. Cell resistance to detachment forces on engineered proteins adsorbed to glass substrates, 2. Western analysis confirming complete cleavage of heptahistidine- and T7-tags, and 3. Cell resistance to detachment forces on engineered proteins with heptahistidine- and T7-tags removed. | ||||||||||||
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Issue or Number: | 1 | ||||||||||||
DOI: | 10.1021/bm049627q | ||||||||||||
Record Number: | CaltechAUTHORS:20150114-121449592 | ||||||||||||
Persistent URL: | https://resolver.caltech.edu/CaltechAUTHORS:20150114-121449592 | ||||||||||||
Official Citation: | Cell-Binding Domain Context Affects Cell Behavior on Engineered Proteins Sarah C. Heilshorn,†, Julie C. Liu,† and, and David A. Tirrell* Biomacromolecules 2005 6 (1), 318-323 | ||||||||||||
Usage Policy: | No commercial reproduction, distribution, display or performance rights in this work are provided. | ||||||||||||
ID Code: | 53712 | ||||||||||||
Collection: | CaltechAUTHORS | ||||||||||||
Deposited By: | Katherine Johnson | ||||||||||||
Deposited On: | 15 Jan 2015 19:06 | ||||||||||||
Last Modified: | 10 Nov 2021 20:06 |
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