PDBsum entry: 2keq

Splicing

PDB id: 2keq

Contents

Protein chain

139 a.a. *

* Residue conservation analysis

PDB id:

2keq

Name:

Splicing

Title:

Solution structure of dnae intein from nostoc punctiforme

Structure:

DNA polymerase iii alpha subunit, nucleic acid binding ob-fold tRNA/helicase-type. Chain: a. Engineered: yes. Mutation: yes. Other_details: fusion protein of n-intein part from DNA polymerase iii alpha subunit (residues 1-102) and c-intein part from nucleic acid binding ob-fold, tRNA/helicase-type (residues 103-137)

Source:

Nostoc punctiforme pcc 73102. Organism_taxid: 63737. Gene: dnae, npun_f4872. Expressed in: escherichia coli. Expression_system_taxid: 562.

NMR struc:

20 models

Authors:

J.S.Oeemig,A.S.Aranko,J.B.Djupsj,H.Iwai

Key ref:

J.S.Oeemig et al. (2009). Solution structure of DnaE intein from Nostoc punctiforme: structural basis for the design of a new split intein suitable for site-specific chemical modification. Febs Lett, 583, 1451-1456. PubMed id: 19344715 DOI: 10.1016/j.febslet.2009.03.058

Date:

02-Feb-09 Release date: 19-May-09

Protein chain

B2J066  (B2J066_NOSP7) -  DNA polymerase III, alpha subunit

Seq: 876 a.a.

Struc: 139 a.a.*

Protein chain

B2J821  (B2J821_NOSP7) -  Nucleic acid binding, OB-fold, tRNA/helicase-type

Seq: 450 a.a.

Struc: 139 a.a.*

* PDB and UniProt seqs differ at 4 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.2.7.7.7 - DNA-directed Dna polymerase.
• Reaction: Deoxynucleoside triphosphate + DNA(n) = diphosphate + DNA(n+1)

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

 Gene Ontology (GO) functional annotation 

• Biological process: intein-mediated protein splicing (1 term)

reference

DOI no: 10.1016/j.febslet.2009.03.058

Febs Lett 583:1451-1456 (2009)

PubMed id: 19344715

Solution structure of DnaE intein from Nostoc punctiforme: structural basis for the design of a new split intein suitable for site-specific chemical modification.

J.S.Oeemig, A.S.Aranko, J.Djupsjöbacka, K.Heinämäki, H.Iwaï.

ABSTRACT

Naturally split DnaE intein from Nostoc punctiforme (Npu) has robust protein trans-splicing activity and high tolerance of sequence variations at the splicing junctions. We determined the solution structure of a single chain variant of NpuDnaE intein by NMR spectroscopy. Based on the NMR structure and the backbone dynamics of the single chain NpuDnaE intein, we designed a functional split variant of the NpuDnaE intein having a short C-terminal half (C-intein) composed of six residues. In vivo and in vitro protein ligation of model proteins by the newly designed split intein were demonstrated.