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PMID: 10097083 Published · ppublish English Comparative Study Journal Article

Crystal structure of a thermostable type B DNA polymerase from Thermococcus gorgonarius.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 96 ·No. 7 ·1999-03-30 ·Pages 3600-5

Hopfner KP, Eichinger A, Engh RA, Laue F, Ankenbauer W, Huber R, Angerer B

Abstract

Most known archaeal DNA polymerases belong to the type B family, which also includes the DNA replication polymerases of eukaryotes, but maintain high fidelity at extreme conditions. We describe here the 2.5 A resolution crystal structure of a DNA polymerase from the Archaea Thermococcus gorgonarius and identify structural features of the fold and the active site that are likely responsible for its thermostable function. Comparison with the mesophilic B type DNA polymerase gp43 of the bacteriophage RB69 highlights thermophilic adaptations, which include the presence of two disulfide bonds and an enhanced electrostatic complementarity at the DNA-protein interface. In contrast to gp43, several loops in the exonuclease and thumb domains are more closely packed; this apparently blocks primer binding to the exonuclease active site. A physiological role of this "closed" conformation is unknown but may represent a polymerase mode, in contrast to an editing mode with an open exonuclease site. This archaeal B DNA polymerase structure provides a starting point for structure-based design of polymerases or ligands with applications in biotechnology and the development of antiviral or anticancer agents.

MeSH Terms
Amino Acid Sequence Binding Sites Cloning, Molecular Computer Graphics Conserved Sequence Crystallography, X-Ray/methods DNA Polymerase I/chemistry,metabolism Enzyme Stability Hot Temperature Models, Molecular Molecular Sequence Data Protein Structure, Secondary Recombinant Proteins/chemistry,metabolism Sequence Alignment Sequence Homology, Amino Acid Thermococcus/enzymology Thermodynamics
Chemicals
Recombinant Proteins DNA Polymerase I
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Hopfner K P
Abteilung Strukturforschung, Max-Planck-Institut für Biochemie, D-82152 Martinsried, Germany. hopfner@scripps.edu
Eichinger A
Engh R A
Laue F
Ankenbauer W
Huber R
Angerer B
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.96.7.3600
Email
hopfner@scripps.edu
Published
1999-03-30
Pages
3600-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC22340
Subset
IM
Databases
PDB
1TGO
External Links
PubMed source DOI: 10.1073/pnas.96.7.3600 PMC Full Text View journal details
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