Reference - Detail
| RRC ID | 5311 |
|---|---|
| Author | Nakayama H, Shimamura T, Imagawa T, Shirai N, Itoh T, Sako Y, Miyano M, Sakuraba H, Ohshima T, Nomura N, Tsuge H. |
| Title | Structure of a hyperthermophilic archaeal homing endonuclease, I-Tsp061I: contribution of cross-domain polar networks to thermostability. |
| Journal | J Mol Biol |
| Abstract |
A novel LAGLIDADG-type homing endonuclease (HEase), I-Tsp061I, from the hyperthermophilic archaeon Thermoproteus sp. IC-061 16 S rRNA gene (rDNA) intron was characterized with respect to its structure, catalytic properties and thermostability. It was found that I-Tsp061I is a HEase isoschizomer of the previously described I-PogI and exhibits the highest thermostability among the known LAGLIDADG-type HEases. Determination of the crystal structure of I-Tsp061I at 2.1 A resolution using the multiple isomorphous replacement and anomalous scattering method revealed that the overall fold is similar to that of other known LAGLIDADG-type HEases, despite little sequence similarity between I-Tsp061I and those HEases. However, I-Tsp061I contains important cross-domain polar networks, unlike its mesophilic counterparts. Notably, the polar network Tyr6-Asp104-His180-107O-HOH12-104O-Asn177 exists across the two packed alpha-helices containing both the LAGLIDADG catalytic motif and the GxxxG hydrophobic helix bundle motif. Another important structural feature is the salt-bridge network Asp29-Arg31-Glu182 across N and C-terminal domain interface, which appears to contribute to the stability of the domain/domain packing. On the basis of these structural analyses and extensive mutational studies, we conclude that such cross-domain polar networks play key roles in stabilizing the catalytic center and domain packing, and underlie the hyperthermostability of I-Tsp061I. |
| Volume | 365(2) |
| Pages | 362-78 |
| Published | 2007-1-12 |
| DOI | 10.1016/j.jmb.2006.09.066 |
| PII | S0022-2836(06)01290-3 |
| PMID | 17069851 |
| MeSH | Amino Acid Motifs Amino Acid Sequence Archaeal Proteins / chemistry* Archaeal Proteins / genetics Binding Sites Crystallization Crystallography, X-Ray Endonucleases / chemistry* Endonucleases / genetics Enzyme Stability Molecular Sequence Data Protein Structure, Tertiary Sequence Homology, Amino Acid Structure-Activity Relationship Thermoproteus / enzymology* |
| IF | 4.76 |
| Times Cited | 9 |
| WOS Category | BIOCHEMISTRY & MOLECULAR BIOLOGY |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 10 |
| Altmetric score changes over past 6months | 3.0 |
| Resource | |
| General Microbes | JCM10595 |