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| IDENTIFICATION OF A TRANSLOCATED PROTEIN SEGMENT IN A VOLTAGE-DEPENDENT CHANNEL |
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| Author(s): SLATIN SL, QIU XQ, JAKES KS, FINKELSTEIN A |
| Source: NATURE Volume: 371 Issue: 6493 Pages: 158-161 Published: SEP 8 1994 |
| Times Cited: 122 References: 19 |
| Abstract: VOLTAGE-GATED channels undergo a conformational change in response to changes in transmembrane voltage. Here we use site-directed biotinylation to create conformation-sensitive sites on colicin Ia, a bacteriocidal protein that forms a voltage-sensitive membrane channel, which can be monitored by electrophysiological methods(1,2). We investigated a model of gating developed for the partly homologous colicin E1 that is based on the insertion of regions of the protein into the membrane in response to cis-positive voltages(3-6). Site-directed cysteine mutagenesis, followed by chemical modification, was used to attach a biotin molecule covalently to a series of unique sites on colicin Ia. The modified protein was incorporated into planar lipid membranes, where the introduced biotin moiety served as a site to bind the water-soluble protein streptavidin, added to one side of the membrane or the other. Our results show that colicin gating is associated with the translocation across the membrane of a segment of the protein of at least 31 amino acids. |
| Document Type: Article |
| Language: English |
| Reprint Address: SLATIN, SL (reprint author), YESHIVA UNIV ALBERT EINSTEIN COLL MED, DEPT PHYSIOL & BIOPHYS, BRONX, NY 10461 USA |
| Publisher: MACMILLAN MAGAZINES LTD, PORTERS SOUTH, 4 CRINAN ST, LONDON, ENGLAND N1 9XW |
| Subject Category: Multidisciplinary Sciences |
| IDS Number: PF191 |
| ISSN: 0028-0836 |
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| | | | | | | | | Record from Web of Science® | | | | | | | | | |