Over-Production, Crystallization, and Preliminary X-ray Crystallographic Analysis of a Coiled-Coil Region in Human Pericentrin
The genes encoding three coiled-coil regions in human pericentrin were gene synthesized with Escherichia coli codon-optimization, and the proteins were successfully over-produced in large quantities using E. coli expression. After verifying that the purified proteins were mostly composed of α-helice...
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MDPI AG
2017-10-01
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| Series: | Crystals |
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| Online Access: | https://www.mdpi.com/2073-4352/7/10/296 |
| _version_ | 1798005557217460224 |
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| author | Min Ye Kim Jeong Kuk Park Yeowon Sim Doheum Kim Jeong Yeon Sim SangYoun Park |
| author_facet | Min Ye Kim Jeong Kuk Park Yeowon Sim Doheum Kim Jeong Yeon Sim SangYoun Park |
| author_sort | Min Ye Kim |
| collection | DOAJ |
| description | The genes encoding three coiled-coil regions in human pericentrin were gene synthesized with Escherichia coli codon-optimization, and the proteins were successfully over-produced in large quantities using E. coli expression. After verifying that the purified proteins were mostly composed of α-helices, one of the proteins was crystallized using polyethylene glycol 8000 as crystallizing agent. X-ray diffraction data were collected to 3.8 Å resolution under cryo-condition using synchrotron X-ray. The crystal belonged to space group C2 with unit cell parameters a = 324.9 Å, b = 35.7 Å, c = 79.5 Å, and β = 101.6˚. According to Matthews’ coefficient, the asymmetric unit may contain up to 12 subunits of the monomeric protein, with a crystal volume per protein mass (VM) of 1.96 Å3 Da−1 and a 37.3% solvent content. |
| first_indexed | 2024-04-11T12:41:04Z |
| format | Article |
| id | doaj.art-8fd571a570ae40388fffd70c74f849d1 |
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| issn | 2073-4352 |
| language | English |
| last_indexed | 2024-04-11T12:41:04Z |
| publishDate | 2017-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Crystals |
| spelling | doaj.art-8fd571a570ae40388fffd70c74f849d12022-12-22T04:23:30ZengMDPI AGCrystals2073-43522017-10-0171029610.3390/cryst7100296cryst7100296Over-Production, Crystallization, and Preliminary X-ray Crystallographic Analysis of a Coiled-Coil Region in Human PericentrinMin Ye Kim0Jeong Kuk Park1Yeowon Sim2Doheum Kim3Jeong Yeon Sim4SangYoun Park5School of Systems Biomedical Science, Soongsil University, Seoul 06978, KoreaSchool of Systems Biomedical Science, Soongsil University, Seoul 06978, KoreaSchool of Systems Biomedical Science, Soongsil University, Seoul 06978, KoreaSchool of Systems Biomedical Science, Soongsil University, Seoul 06978, KoreaSchool of Systems Biomedical Science, Soongsil University, Seoul 06978, KoreaSchool of Systems Biomedical Science, Soongsil University, Seoul 06978, KoreaThe genes encoding three coiled-coil regions in human pericentrin were gene synthesized with Escherichia coli codon-optimization, and the proteins were successfully over-produced in large quantities using E. coli expression. After verifying that the purified proteins were mostly composed of α-helices, one of the proteins was crystallized using polyethylene glycol 8000 as crystallizing agent. X-ray diffraction data were collected to 3.8 Å resolution under cryo-condition using synchrotron X-ray. The crystal belonged to space group C2 with unit cell parameters a = 324.9 Å, b = 35.7 Å, c = 79.5 Å, and β = 101.6˚. According to Matthews’ coefficient, the asymmetric unit may contain up to 12 subunits of the monomeric protein, with a crystal volume per protein mass (VM) of 1.96 Å3 Da−1 and a 37.3% solvent content.https://www.mdpi.com/2073-4352/7/10/296pericentrincoiled-coilcentrosomepericentriolar material (PCM) |
| spellingShingle | Min Ye Kim Jeong Kuk Park Yeowon Sim Doheum Kim Jeong Yeon Sim SangYoun Park Over-Production, Crystallization, and Preliminary X-ray Crystallographic Analysis of a Coiled-Coil Region in Human Pericentrin Crystals pericentrin coiled-coil centrosome pericentriolar material (PCM) |
| title | Over-Production, Crystallization, and Preliminary X-ray Crystallographic Analysis of a Coiled-Coil Region in Human Pericentrin |
| title_full | Over-Production, Crystallization, and Preliminary X-ray Crystallographic Analysis of a Coiled-Coil Region in Human Pericentrin |
| title_fullStr | Over-Production, Crystallization, and Preliminary X-ray Crystallographic Analysis of a Coiled-Coil Region in Human Pericentrin |
| title_full_unstemmed | Over-Production, Crystallization, and Preliminary X-ray Crystallographic Analysis of a Coiled-Coil Region in Human Pericentrin |
| title_short | Over-Production, Crystallization, and Preliminary X-ray Crystallographic Analysis of a Coiled-Coil Region in Human Pericentrin |
| title_sort | over production crystallization and preliminary x ray crystallographic analysis of a coiled coil region in human pericentrin |
| topic | pericentrin coiled-coil centrosome pericentriolar material (PCM) |
| url | https://www.mdpi.com/2073-4352/7/10/296 |
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