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| MEMBRANE TOPOLOGY OF ESCHERICHIA-COLI DIACYLGLYCEROL KINASE |
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| Author(s): SMITH RL, OTOOLE JF, MAGUIRE ME, SANDERS CR |
| Source: JOURNAL OF BACTERIOLOGY Volume: 176 Issue: 17 Pages: 5459-5465 Published: SEP 1994 |
| Times Cited: 43 References: 37 |
| Abstract: The topology of Escherichia coli diacylglycerol kinase (DAGK) within the cytoplasmic membrane was elucidated by a combined approach involving both multiple aligned sequence analysis and fusion protein experiments. Hydropathy plots of the five prokaryotic DAGK sequences available were uniform in their prediction of three transmembrane segments. The hydropathy predictions were experimentally tested genetically by fusing C-terminal deletion derivatives of DAGK to beta-lactamase and beta-galactosidase. Following expression, the enzymatic activities of the chimeric proteins were measured and used to determine the cellular location of the fusion junction. These studies confirmed the hydropathy predictions for DAGK with respect to the number and approximate sequence locations of the transmembrane segments. Further analysis of the aligned DAGK sequences detected probable ex-helical N-terminal capping motifs and two amphipathic alpha-helices within the enzyme. The combined fusion and sequence data indicate that DAGK is a polytopic integral membrane protein with three transmembrane segments with the N terminus of the protein in the cytoplasm, the C terminus in the periplasmic space, and two amphipathic helices near the cytoplasmic surface. |
| Document Type: Article |
| Language: English |
Addresses:
1. CASE WESTERN RESERVE UNIV, SCH MED, DEPT PHYSIOL & BIOPHYS, CLEVELAND, OH 44106 USA
2. CASE WESTERN RESERVE UNIV, SCH MED, DEPT PHARMACOL, CLEVELAND, OH 44106 USA |
| Publisher: AMER SOC MICROBIOLOGY, 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 |
| Subject Category: Microbiology |
| IDS Number: PD457 |
| ISSN: 0021-9193 |
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| | | | | | | | | Record from Web of Science® | | | | | | | | | |