Beissinger, Martina and Paulus, Christina and Bayer, Peter and Wolf, Hans and Rösch, Paul and Wagner, Ralf (1996) Sequence-specific resonance assignments of the H-1-NMR spectra and structural characterization in solution of the HIV-1 transframe protein p6. EUROPEAN JOURNAL OF BIOCHEMISTRY, 237 (2). pp. 383-392. ISSN 0014-2956
Full text not available from this repository.Abstract
The frameshift protein p6* encoded directly upstream of the protease in the human immunodeficiency virus type 1 (HIV-1) pol reading frame is thought to be a natural inhibitor of protease activation and to play a role in the polyprotein processing of Gag and Gag-Pol precursors. To allow structural characterization of the p6* transframe protein, the p6* coding region was cloned into the vector pGEX-KG and expressed in Escherichia coli as a fusion protein with glutathione S-transferase (GST) under the control of the tac promoter. Thrombin cleavage of the construct resulted in a 70-amino-acid polypeptide which is extended by two additional residues at the N-terminus compared to the natural p6* sequence. The native purification procedure including an affinity and a size-exclusion chromatography step yielded sufficient amounts of highly pure protein suitable for NMR spectroscopy. Fluorescence, circular dichroism and H-1-NMR spectroscopy were applied to characterize the structure of the protein. Two-dimensional NMR spectra provided essentially complete sequence-specific resonance assignments at pH 5.9. Although there is evidence for a helix-forming tendency in the N-terminus of the protein, the experiments indicate that p6* has no overall stable secondary or tertiary structure with the single tryptophan exposed in aqueous solution. However, the results reported herein open the way to characterize further the interaction of p6* with the HIV-1 protease in structural and functional in vitro studies.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | SECONDARY STRUCTURE PREDICTION; IMMUNODEFICIENCY-VIRUS TYPE-1; INTERFACE PEPTIDES; VIRAL INFECTIVITY; ESCHERICHIA-COLI; GENE-PRODUCT; INHIBITION; PROTEASES; IDENTIFICATION; SPECTROSCOPY; H-1-NMR; human immunodeficiency virus 1; protease regulation; protein purification; transframe protein p6 |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Medizinische Mikrobiologie und Hygiene |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 02 Nov 2023 07:06 |
| Last Modified: | 02 Nov 2023 07:06 |
| URI: | https://pred.uni-regensburg.de/id/eprint/51797 |
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