Identification, Characterization, and Preliminary X-ray Diffraction Analysis of a Single Stranded DNA Binding Protein (LjSSB) from Psychrophilic <i>Lacinutrix jangbogonensis</i> PAMC 27137
Single-stranded DNA-binding proteins (SSBs) are essential for DNA metabolism, including repair and replication, in all organisms. SSBs have potential applications in molecular biology and in analytical methods. In this study, for the first time, we purified, structurally characterized, and analyzed...
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MDPI AG
2022-04-01
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| author | Woong Choi Jonghyeon Son Aekyung Park Hongshi Jin Seung Chul Shin Jun Hyuck Lee T. Doohun Kim Han-Woo Kim |
| author_facet | Woong Choi Jonghyeon Son Aekyung Park Hongshi Jin Seung Chul Shin Jun Hyuck Lee T. Doohun Kim Han-Woo Kim |
| author_sort | Woong Choi |
| collection | DOAJ |
| description | Single-stranded DNA-binding proteins (SSBs) are essential for DNA metabolism, including repair and replication, in all organisms. SSBs have potential applications in molecular biology and in analytical methods. In this study, for the first time, we purified, structurally characterized, and analyzed psychrophilic SSB (LjSSB) from <i>Lacinutrix jangbogonensis</i> PAMC 27137 isolated from the Antarctic region. LjSSB has a relatively short amino acid sequence, consisting of 111 residues, with a molecular mass of 12.6 kDa. LjSSB protein was overexpressed in <i>Escherichia coli</i> BL21 (DE3) and analyzed for binding affinity using 20- and 35-mer deoxythymidine oligonucleotides (dT). In addition, the crystal structure of LjSSB at a resolution 2.6 Å was obtained. The LjSSB protein crystal belongs to the space group <i>C</i>222 with the unit cell parameters of a = 106.58 Å, b = 234.14 Å, c = 66.14 Å. The crystal structure was solved using molecular replacement, and subsequent iterative structure refinements and model building are currently under progress. Further, the complete structural information of LjSSB will provide a novel strategy for protein engineering and for the application on molecular biological techniques. |
| first_indexed | 2024-03-09T10:59:24Z |
| format | Article |
| id | doaj.art-ac7ce4be027f41c6bb1493964cbec6c5 |
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| issn | 2073-4352 |
| language | English |
| last_indexed | 2024-03-09T10:59:24Z |
| publishDate | 2022-04-01 |
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| spelling | doaj.art-ac7ce4be027f41c6bb1493964cbec6c52023-12-01T01:25:38ZengMDPI AGCrystals2073-43522022-04-0112453810.3390/cryst12040538Identification, Characterization, and Preliminary X-ray Diffraction Analysis of a Single Stranded DNA Binding Protein (LjSSB) from Psychrophilic <i>Lacinutrix jangbogonensis</i> PAMC 27137Woong Choi0Jonghyeon Son1Aekyung Park2Hongshi Jin3Seung Chul Shin4Jun Hyuck Lee5T. Doohun Kim6Han-Woo Kim7Research Unit of Cryogenic Novel Material, Korea Polar Research Institute, Incheon 21990, KoreaNew Drug Development Center, Daegu-Gyeongbuk Medical Innovation Foundation, Daegu 41061, KoreaDivision of Emerging Infectious Diseases Bureau of Infectious Disease Diagnosis Control, Korea Disease Control and Prevention Agency, Cheongju 28159, KoreaDivision of Life Sciences, Korea Polar Research Institute, Incheon 21990, KoreaDivision of Life Sciences, Korea Polar Research Institute, Incheon 21990, KoreaResearch Unit of Cryogenic Novel Material, Korea Polar Research Institute, Incheon 21990, KoreaDepartment of Chemistry, Sookmyung Women’s University, Seoul 04310, KoreaResearch Unit of Cryogenic Novel Material, Korea Polar Research Institute, Incheon 21990, KoreaSingle-stranded DNA-binding proteins (SSBs) are essential for DNA metabolism, including repair and replication, in all organisms. SSBs have potential applications in molecular biology and in analytical methods. In this study, for the first time, we purified, structurally characterized, and analyzed psychrophilic SSB (LjSSB) from <i>Lacinutrix jangbogonensis</i> PAMC 27137 isolated from the Antarctic region. LjSSB has a relatively short amino acid sequence, consisting of 111 residues, with a molecular mass of 12.6 kDa. LjSSB protein was overexpressed in <i>Escherichia coli</i> BL21 (DE3) and analyzed for binding affinity using 20- and 35-mer deoxythymidine oligonucleotides (dT). In addition, the crystal structure of LjSSB at a resolution 2.6 Å was obtained. The LjSSB protein crystal belongs to the space group <i>C</i>222 with the unit cell parameters of a = 106.58 Å, b = 234.14 Å, c = 66.14 Å. The crystal structure was solved using molecular replacement, and subsequent iterative structure refinements and model building are currently under progress. Further, the complete structural information of LjSSB will provide a novel strategy for protein engineering and for the application on molecular biological techniques.https://www.mdpi.com/2073-4352/12/4/538single stranded DNA binding protein<i>Lacinutrix jangbogonensis</i> PAMC 27137X-ray crystallography |
| spellingShingle | Woong Choi Jonghyeon Son Aekyung Park Hongshi Jin Seung Chul Shin Jun Hyuck Lee T. Doohun Kim Han-Woo Kim Identification, Characterization, and Preliminary X-ray Diffraction Analysis of a Single Stranded DNA Binding Protein (LjSSB) from Psychrophilic <i>Lacinutrix jangbogonensis</i> PAMC 27137 Crystals single stranded DNA binding protein <i>Lacinutrix jangbogonensis</i> PAMC 27137 X-ray crystallography |
| title | Identification, Characterization, and Preliminary X-ray Diffraction Analysis of a Single Stranded DNA Binding Protein (LjSSB) from Psychrophilic <i>Lacinutrix jangbogonensis</i> PAMC 27137 |
| title_full | Identification, Characterization, and Preliminary X-ray Diffraction Analysis of a Single Stranded DNA Binding Protein (LjSSB) from Psychrophilic <i>Lacinutrix jangbogonensis</i> PAMC 27137 |
| title_fullStr | Identification, Characterization, and Preliminary X-ray Diffraction Analysis of a Single Stranded DNA Binding Protein (LjSSB) from Psychrophilic <i>Lacinutrix jangbogonensis</i> PAMC 27137 |
| title_full_unstemmed | Identification, Characterization, and Preliminary X-ray Diffraction Analysis of a Single Stranded DNA Binding Protein (LjSSB) from Psychrophilic <i>Lacinutrix jangbogonensis</i> PAMC 27137 |
| title_short | Identification, Characterization, and Preliminary X-ray Diffraction Analysis of a Single Stranded DNA Binding Protein (LjSSB) from Psychrophilic <i>Lacinutrix jangbogonensis</i> PAMC 27137 |
| title_sort | identification characterization and preliminary x ray diffraction analysis of a single stranded dna binding protein ljssb from psychrophilic i lacinutrix jangbogonensis i pamc 27137 |
| topic | single stranded DNA binding protein <i>Lacinutrix jangbogonensis</i> PAMC 27137 X-ray crystallography |
| url | https://www.mdpi.com/2073-4352/12/4/538 |
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