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The gram-positive bacterium Staphylococcus aureus is a common cause of abscesses, sinusitis and food poisoning. Theemergence of antibiotic-resistant strains has caused significant clinical issues worldwide. The HslU-HslV complex wasfirst identified as a prokaryotic homolog of eukaryotic proteasomes. HslU is an unfoldase that mediates the unfolding ofthe substrate proteins, and it works with the protease HslV in the complex. To date, the protein complex has been mostlystudied in gram-negative bacteria. In this study, we report the purification and crystallization of the full-length HslU fromS. aureus. The crystal diffracted X-rays to a 3.5 Å resolution, revealing that the crystals belong to space group P21, withunit cell parameters of a = 166.5, b = 189.6, c = 226.6 Å, and β = 108.1°. We solved the phage problem by molecularreplacement using the structure of HslU from Haemophilus influenzae as a search model. The cell content analysis with thismolecular replacement solution revealed that 24 molecules are contained in the asymmetric unit. This structure providesinsight into the structural and mechanistic difference of the HslUV complex of gram-positive bacteria.
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참고문헌 (9)
참고문헌 신청
Cabrita, L. D / 2006 / A family of E. coli expression vectors for laboratory scale and high throughput soluble protein production / BMC Biotechnol 6:12
Goldberg, A. L / 1976 / Intracellular protein degradation in mammalian and bacterial cells : Part 2 / Annu Rev Biochem 45:747~803
Knecht, E / 2009 / Intracellular protein degradation in mammalian cells : recent developments / Cell Mol Life Sci 66:2427~2443
Kwon, A. R / 2003 / Structure and reactivity of an asymmetric complex between HslV and I-domain deleted HslU, a prokaryotic homolog of the eukaryotic proteasome / J Mol Biol 330:185~195
Otwinowski, Z / 1997 / Processing of X-ray diffraction data collected in oscillation mode / Methods Enzymol 276:307~326
Goldberg, A. L / 1976 / Intracellular protein degradation in mammalian and bacterial cells : Part 2 / Annu Rev Biochem 45:747~803
Knecht, E / 2009 / Intracellular protein degradation in mammalian cells : recent developments / Cell Mol Life Sci 66:2427~2443
Kwon, A. R / 2003 / Structure and reactivity of an asymmetric complex between HslV and I-domain deleted HslU, a prokaryotic homolog of the eukaryotic proteasome / J Mol Biol 330:185~195
Otwinowski, Z / 1997 / Processing of X-ray diffraction data collected in oscillation mode / Methods Enzymol 276:307~326
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