The asymmetric unit contains a dimer of the NS3-NS4a complex. This is only the protease domain of NS3 and a peptide of NS4a. This dimeric structure is the result of the soultion structure of the domain. The full length NS3 contains a 3-domain helicase as well and would not have the same dimeric interfaces.
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Components
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Protein / Protein/peptide , 2 types, 4 molecules ACBD
#1: Protein
HCVNS3Protease
Mass: 21233.225 Da / Num. of mol.: 2 / Fragment: Protease domain, UNP residues 1027-1207 Source method: isolated from a genetically manipulated source Details: T7 epitope (MASMTGGQQMG)followed by HCV NS3 residues 1-181 and the His-tag (SGHHHHHH) Source: (gene. exp.) Hepatitis C virus subtype 1a / Strain: H77 Strain of genotype 1a / Gene: NS3 / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3) / References: UniProt: Q9ELS8
#2: Protein/peptide
HCVNS4apeptide
Mass: 2394.039 Da / Num. of mol.: 2 / Fragment: UNP residues 1678-1696 / Mutation: C28S / Source method: obtained synthetically Details: Peptides were synthesized using Fmoc solid-phase chemistry on an ABI 431 synthesizer (Foster City, CA). Preloaded 2-chlorotrityl chloride resin or Wang resin was used for the solid phase ...Details: Peptides were synthesized using Fmoc solid-phase chemistry on an ABI 431 synthesizer (Foster City, CA). Preloaded 2-chlorotrityl chloride resin or Wang resin was used for the solid phase assembly of NS4A activator peptide. The sequence of the peptide was Lys-Lys-Gly-Ser-Val-Val-Ile-Val-Gly-Arg-Ile-Ile-Leu-Ser-Gly-Arg-Pro-Ala-Ile-Val-Pro-Lys-Lys-OH. Trifunctional residue sidechain protecting groups included tert-butyl for Ser, tert-butoxycarbonyl for Lys, and 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl for Arg. Cleavage and sidechain deprotection was accomplished using 92.5% trifluoroacetic acid, with 2.5% each of water, ethanedithiol and triisopropylsilane for 2 hours. The peptide was purified by reversed phase HPLC. The peptide molecular weight was confirmed by electrospray ionization mass spectrometry. References: UniProt: Q9ELS8
Mass: 18.015 Da / Num. of mol.: 164 / Source method: isolated from a natural source / Formula: H2O
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Experimental details
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Experiment
Experiment
Method: X-RAY DIFFRACTION / Number of used crystals: 1
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Sample preparation
Crystal
Density Matthews: 3.83 Å3/Da / Density % sol: 67.9 %
Crystal grow
Temperature: 285 K / Method: vapor diffusion, hanging drop / pH: 5.6 Details: Crystallization was performed by the vapor diffusion method using hanging drops (4 L protein solution mixed with 4 L (0.75 - 1.00) M NaCl - 0.1 M MES - 0.1 M Na/K PO4, pH 5.6 - 6.2) ...Details: Crystallization was performed by the vapor diffusion method using hanging drops (4 L protein solution mixed with 4 L (0.75 - 1.00) M NaCl - 0.1 M MES - 0.1 M Na/K PO4, pH 5.6 - 6.2) suspended over 1 mL reservoir solutions of (1.25 - 1.50) M NaCl - 0.1 M MES - 0.1 M Na/K PO4 - 5 mM -mercaptoethanol, pH 5.6-6.2. The trays were set at 4oC for 5-7 days to control nucleation, followed by incubation for 3 weeks at 12oC to maximize crystal growth. , VAPOR DIFFUSION, HANGING DROP, temperature 277, then 285K
In the structure databanks used in Yorodumi, some data are registered as the other names, "COVID-19 virus" and "2019-nCoV". Here are the details of the virus and the list of structure data.
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