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Prevention of Aggregation of Synthetic Membrane-Spanning Peptides by Addition of Detergent

Tomich, John M. and Carson, L. Wulf and Kanes, Katherine J. and Vogelaar, Nancy J. and Emerling, Michael R. and Richards, John H. (1988) Prevention of Aggregation of Synthetic Membrane-Spanning Peptides by Addition of Detergent. Analytical Biochemistry, 174 (1). pp. 197-203. ISSN 0003-2697. http://resolver.caltech.edu/CaltechAUTHORS:20150428-074254869

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Abstract

In our initial attempts to solubilize and purify a chemically synthesized 22-amino acid, membrane-spanning peptide, we encountered numerous difficulties. The peptide was not soluble in dilute acids, organic solvents, or chaotropic agents (±detergent) following standard HF cleavage protocols. The insolubility was a direct result of the formation of peptide-(peptide)_n aggregates that occurred during the initial phase of isolation, i.e., during the HF cleavage. Eliminating the ether precipitation and subsequent washes did not decrease the degree of aggregation of the product. Inclusion of nonionic detergents in the HF-cleavage reactions displayed little ability in preventing aggregation. Cleavage in the presence of sodium dodecyl sulfate, however, dramatically reduced the degree of aggregation, even after washing with organic solvents. The cleaved peptide was purified to homogeneity using a detergent-based HPLC protocol. This column procedure also permits the quantitative exchange of the sodium dodecyl sulfate for n-octyl-β-d-glucopyranoside. Combined use of the two protocols results in high-yield isolations for a class of peptides that is generally difficult to handle.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1016/0003-2697(88)90535-0DOIArticle
http://www.sciencedirect.com/science/article/pii/0003269788905350PublisherArticle
Additional Information:© 1988 Academic Press, Inc. Received 28 March 1988. The authors to thank Professors John A. Barranger, Mauricio Montal, and Julian C. Williams for helpful discussions during preparation of the manuscript and Elizabeth Aguirre for preparation of the manuscript.
Subject Keywords: detergent; peptide synthesis; HPLC; peptide purification; membrane peptides
Record Number:CaltechAUTHORS:20150428-074254869
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20150428-074254869
Official Citation:John M. Tomich, L.Wulf Carson, Katherine J. Kanes, Nancy J. Vogelaar, Michael R. Emerling, John H. Richards, Prevention of aggregation of synthetic membrane-spanning peptides by addition of detergent, Analytical Biochemistry, Volume 174, Issue 1, October 1988, Pages 197-203, ISSN 0003-2697, http://dx.doi.org/10.1016/0003-2697(88)90535-0. (http://www.sciencedirect.com/science/article/pii/0003269788905350)
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ID Code:57032
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:28 Apr 2015 19:14
Last Modified:28 Apr 2015 19:14

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