PDBsum entry 4pls

Peptide binding protein

PDB id: 4pls

Contents

Protein chains

280 a.a.

Ligands

ACT ×2

Metals

_CA ×10

Waters ×210

PDB id:

4pls

Name:

Peptide binding protein

Title:

Crystal structures of designed armadillo repeat proteins: implications of construct design and crystallization conditions on overall structure.

Structure:

Arm00010. Chain: a, b, c, d. Engineered: yes. Other_details: artificial sequence

Source:

Synthetic construct. Organism_taxid: 32630. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

2.35Å R-factor: 0.187 R-free: 0.267

Authors:

C.Reichen,C.Madhurantakam,A.Plueckthun,P.R.Mittl

Key ref:

C.Reichen et al. (2014). Crystal structures of designed armadillo repeat proteins: implications of construct design and crystallization conditions on overall structure. Protein Sci, 23, 1572-1583. PubMed id: 25132085 DOI: 10.1002/pro.2535

Date:

19-May-14 Release date: 10-Dec-14

Protein chains

No UniProt id for this chain

Struc: 280 a.a.

DOI no: 10.1002/pro.2535

Protein Sci 23:1572-1583 (2014)

PubMed id: 25132085

Crystal structures of designed armadillo repeat proteins: implications of construct design and crystallization conditions on overall structure.

C.Reichen, C.Madhurantakam, A.Plückthun, P.R.Mittl.

ABSTRACT

Designed armadillo repeat proteins (dArmRP) are promising modular proteins for the engineering of binding molecules that recognize extended polypeptide chains. We determined the structure of a dArmRP containing five internal repeats and 3rd generation capping repeats in three different states by X-ray crystallography: without N-terminal His6 -tag and in the presence of calcium (YM5 A/Ca(2+) ), without N-terminal His6 -tag and in the absence of calcium (YM5 A), and with N-terminal His6 -tag and in the presence of calcium (His-YM5 A/Ca(2+) ). All structures show different quaternary structures and superhelical parameters. His-YM5 A/Ca(2+) forms a crystallographic dimer, which is bridged by the His6 -tag, YM5 A/Ca(2+) forms a domain-swapped tetramer, and only in the absence of calcium and the His6 -tag, YM5 A forms a monomer. The changes of superhelical parameters are a consequence of calcium binding, because calcium ions interact with negatively charged residues, which can also participate in the modulation of helix dipole moments between adjacent repeats. These observations are important for further optimizations of dArmRPs and provide a general illustration of how construct design and crystallization conditions can influence the exact structure of the investigated protein.