Hewick, Rodney M. and Hunkapiller, Michael W. and Hood, Leroy E. and Dreyer, William J. (1981) A gas-liquid solid phase peptide and protein sequenator. Journal of Biological Chemistry, 256 (15). pp. 7990-7997. ISSN 0021-9258. http://resolver.caltech.edu/CaltechAUTHORS:HEWjbc81
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A new miniaturized protein and peptide sequenator has been constructed which uses gas phase reagents at the coupling and cleavage steps of the Edman degradation. The sample is embedded in a matrix of Polybrene dried onto a porous glass fiber disc located in a small cartridge-style reaction cell. The protein or peptide, though not covalently attached to the support, is essentially immobile throughout the degradative cycle, since only relatively apolar, liquid phase solvents pass through the cell. This instrument can give useful sequence data on as little as 5 pmol or protein, can perform extended sequence runs (greater than 30 residues) on subnanomole quantities of proteins purified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and can sequence hydrophobic peptides to completion. The sequenator is characterized by a high repetitive yield during the degradation, low reagent consumption, low maintenance requirements, and a degradative cycle time of only 50 min using a complete double cleavage program.
|Additional Information:||© 1981 by the American Society for Biochemistry and Molecular Biology. Received for publication, April 23, 1981. This work was supported by the Weingart Foundation, National Science Foundation Grant PCM 80-05599, and National Institutes of Health Grant GM 06965. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. We gratefully acknowledge the earlier contributions of David Helphrey and Dr. Suzanna J. Horvath (Division of Biology, California Institute of Technology) toward the general development of the gas-liquid phase system. We are indebted to Anton Stark and other members of the Divisions of Chemistry, Biology, and Central Engineering workshops at the California Institute of Technology who assisted in the design and fabrication of components of the sequenator. We thank also Chin Sook Kim (Divison of Biology, California Institute of Technology) for meticulous purification of various sequenator reagents and solvents.|
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|Deposited By:||Tony Diaz|
|Deposited On:||17 Sep 2008 04:59|
|Last Modified:||26 Dec 2012 10:17|
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