5SZT

Crystal structure of the large fragment of DNA Polymerase I from Thermus aquaticus in a closed ternary complex with 7-(N-(10-hydroxydecanoyl)-aminopentenyl)-7-deaza-2'-dATP

PDB DOI: https://doi.org/10.2210/pdb5SZT/pdb
NAKB: 5SZT

Classification: TRANSFERASE
Organism(s): Thermus aquaticus, synthetic construct
Expression System: Escherichia coli
Mutation(s): No

Deposited: 2016-08-15 Released: 2016-12-14
Deposition Author(s): Hottin, A., Betz, K., Marx, A.

Experimental Data Snapshot

Method: X-RAY DIFFRACTION
Resolution: 1.80 Å
R-Value Free: 0.230
R-Value Work: 0.194
R-Value Observed: 0.195

wwPDB Validation 3D Report Full Report

Ligand Structure Quality Assessment

This is version 1.3 of the entry. See complete history.

Literature

Structural Basis for the KlenTaq DNA Polymerase Catalysed Incorporation of Alkene- versus Alkyne-Modified Nucleotides.

Hottin, A., Betz, K., Diederichs, K., Marx, A.

(2017) Chemistry 23: 2109-2118

PubMed: 27901305 Search on PubMed
DOI: https://doi.org/10.1002/chem.201604515
Primary Citation of Related Structures:
5E41, 5SZT

PubMed Abstract:
Efficient incorporation of modified nucleotides by DNA polymerases is essential for many cutting-edge biomolecular technologies. The present study compares the acceptance of either alkene- or alkyne-modified nucleotides by KlenTaq DNA polymerase and provides structural insights into how 7-deaza-adenosine and deoxyuridine with attached alkene-modifications are incorporated into the growing DNA strand. Thereby, we identified modified nucleotides that prove to be superior substrates for KlenTaq DNA polymerase compared with their natural analogues. The knowledge can be used to guide future design of functionalized nucleotide building blocks.

Organizational Affiliation

Department of Chemistry and Department of Biology, University of Konstanz, Universitätsstrasse 10, 78457, Konstanz, Germany.

Macromolecule Content

Total Structure Weight: 70.73 kDa
Atom Count: 5,309
Modelled Residue Count: 568
Deposited Residue Count: 568
Unique protein chains: 1
Unique nucleic acid chains: 2

Macromolecules

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(by identity cutoff) | 3D Structure

Entity ID: 1
Molecule	Chains	Sequence Length	Organism	Details	Image
DNA polymerase I, thermostable	A	540	Thermus aquaticus	Mutation(s): 0 Gene Names: polA, pol1 EC: 2.7.7.7
UniProt
Find proteins for P19821 (Thermus aquaticus) Explore P19821 Go to UniProtKB: P19821
Entity Groups
Sequence Clusters	30% Identity50% Identity70% Identity90% Identity95% Identity100% Identity
UniProt Group	P19821
Sequence Annotations Expand
Reference Sequence

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Entity ID: 2
Molecule	Chains	Length	Organism	Image
DNA (5'-D(GPAPCPCPAPCPGPGPCPGPCP(DOC))-3')	B	12	synthetic construct
Sequence Annotations Expand
Reference Sequence

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Entity ID: 3
Molecule	Chains	Length	Organism	Image
DNA (5'-D(APAPAPTPGPGPCPGPCPCPGPTPGPGPTPC)-3')	C	16	synthetic construct
Sequence Annotations Expand
Reference Sequence

Small Molecules

Ligands 4 Unique
ID	Chains	Name / Formula / InChI Key	2D Diagram	3D Interactions
72J Query on 72J Download Ideal Coordinates CCD File SDF format, chain J [auth A] MOL2 format, chain J [auth A]	J [auth A]	7-(N-(10-hydroxydecanoyl)-aminopentenyl)-7-deaza-2'-dATP C₂₆ H₄₄ N₅ O₁₄ P₃ KGSINQKQGHRQNT-MNMHHJAXSA-N		Interactions Focus chain J [auth A] Interactions & Density Focus chain J [auth A]
GOL Query on GOL Download Ideal Coordinates CCD File SDF format, chain L [auth C] SDF format, chain F [auth A] MOL2 format, chain L [auth C] MOL2 format, chain F [auth A]	F [auth A], L [auth C]	GLYCEROL C₃ H₈ O₃ PEDCQBHIVMGVHV-UHFFFAOYSA-N		Interactions Focus chain F [auth A] Focus chain L [auth C] Interactions & Density Focus chain F [auth A] Focus chain L [auth C]
ACT Query on ACT Download Ideal Coordinates CCD File SDF format, chain G [auth A] SDF format, chain H [auth A] SDF format, chain I [auth A] SDF format, chain M [auth C] MOL2 format, chain G [auth A] MOL2 format, chain H [auth A] MOL2 format, chain I [auth A] MOL2 format, chain M [auth C]	G [auth A], H [auth A], I [auth A], M [auth C]	ACETATE ION C₂ H₃ O₂ QTBSBXVTEAMEQO-UHFFFAOYSA-M		Interactions Focus chain G [auth A] Focus chain H [auth A] Focus chain I [auth A] Focus chain M [auth C] Interactions & Density Focus chain G [auth A] Focus chain H [auth A] Focus chain I [auth A] Focus chain M [auth C]
MG Query on MG Download Ideal Coordinates CCD File SDF format, chain K [auth B] SDF format, chain D [auth A] SDF format, chain E [auth A] MOL2 format, chain K [auth B] MOL2 format, chain D [auth A] MOL2 format, chain E [auth A]	D [auth A], E [auth A], K [auth B]	MAGNESIUM ION Mg JLVVSXFLKOJNIY-UHFFFAOYSA-N		Interactions Focus chain D [auth A] Focus chain E [auth A] Focus chain K [auth B] Interactions & Density Focus chain D [auth A] Focus chain E [auth A] Focus chain K [auth B]

Experimental Data & Validation

Experimental Data

Method: X-RAY DIFFRACTION
Resolution: 1.80 Å
R-Value Free: 0.230
R-Value Work: 0.194
R-Value Observed: 0.195
Space Group: P 3₁ 2 1

Unit Cell:

Length ( Å )	Angle ( ˚ )
a = 108.425	α = 90
b = 108.425	β = 90
c = 90.615	γ = 120

Software Package:

Software Name	Purpose
PHENIX	refinement
XDS	data reduction
Coot	model building

Structure Validation

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Ligand Structure Quality Assessment

Entry History

Deposition Data

Released Date: 2016-12-14

Deposition Author(s):

Hottin, A.

Betz, K.

Marx, A.

Revision History (Full details and data files)

Version 1.0: 2016-12-14
Type: Initial release
Version 1.1: 2017-02-22
Changes: Database references
Version 1.2: 2019-10-16
Changes: Advisory, Data collection
Version 1.3: 2024-01-17
Changes: Advisory, Data collection, Database references, Derived calculations, Refinement description