PDBsum entry 4zsx

Unknown function

PDB id: 4zsx

Contents

Protein chains

293 a.a.

Ligands

PO4 ×4

Waters ×76

PDB id:

4zsx

Name:

Unknown function

Title:

Structure of a fusion protein with a helix linker, 2arh-3-3kaw-2.0

Structure:

Uncharacterized fusion protein. Chain: a, b. Engineered: yes. Mutation: yes

Source:

Aquifex aeolicus, pseudomonas aeruginosa. Organism_taxid: 224324, 208964. Strain: vf5, atcc 15692 / pao1 / 1c / prs 101 / lmg 12228. Gene: aq_1966, pa2107. Expressed in: escherichia coli. Expression_system_taxid: 469008.

Resolution:

2.19Å R-factor: 0.229 R-free: 0.249

Authors:

Y.-T.Lai,T.O.Yeates

Key ref:

Y.T.Lai et al. (2015). On the predictability of the orientation of protein domains joined by a spanning alpha-helical linker. Protein Eng Des Sel, 28, 491-499. PubMed id: 26243886 DOI: 10.1093/protein/gzv035

Date:

14-May-15 Release date: 19-Aug-15

Protein chains

O67778  (O67778_AQUAE) -  DUF1122 domain-containing protein from Aquifex aeolicus (strain VF5)

Seq: 201 a.a.

Struc: 293 a.a.*

Protein chains

Q9I208  (Q9I208_PSEAE) -  Four-helix bundle copper-binding protein from Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)

Seq: 132 a.a.

Struc: 293 a.a.*

* PDB and UniProt seqs differ at 46 residue positions (black crosses)

DOI no: 10.1093/protein/gzv035

Protein Eng Des Sel 28:491-499 (2015)

PubMed id: 26243886

On the predictability of the orientation of protein domains joined by a spanning alpha-helical linker.

Y.T.Lai, L.Jiang, W.Chen, T.O.Yeates.

ABSTRACT

Connecting proteins together in prescribed geometric arrangements is an important element in new areas of biomolecular design. In this study, we characterize the degree of three-dimensional orientational control that can be achieved when two protein domains that have alpha-helical termini are joined using an alpha-helical linker. A fusion between naturally oligomeric protein domains was designed in this fashion with the intent of creating a self-assembling 12-subunit tetrahedral protein cage. While the designed fusion protein failed to assemble into a tetrahedral cage in high yield, a series of crystal structures showed that the two fused components were indeed bridged by an intact alpha helix, although the fusion protein was distorted from the intended ideal configuration by bending of the helix, ranging from 7 to 35°. That range of deviation in orientation creates challenges for designing large, perfectly symmetric protein assemblies, although it should offer useful outcomes for other less geometrically demanding applications in synthetic biology.