PDBsum entry 5lec

De novo protein

PDB id: 5lec

Contents

Protein chain

474 a.a.

Ligands

B3P

Waters ×9

PDB id:

5lec

Name:

De novo protein

Title:

Crystal structure of darpin-darpin rigid fusion, variant ddd_d12_12_d12_12_d12

Structure:

Ddd_d12_12_d12_12_d12. Chain: a. Engineered: yes

Source:

Synthetic construct. Organism_taxid: 32630. Expressed in: escherichia coli k-12. Expression_system_taxid: 83333. Expression_system_variant: xl1-blue

Resolution:

2.51Å R-factor: 0.234 R-free: 0.266

Authors:

A.Batyuk,Y.Wu,P.R.Mittl,A.Plueckthun

Key ref:

Y.Wu et al. (2017). Rigidly connected multispecific artificial binders with adjustable geometries. Sci Rep, 7, 11217. PubMed id: 28894181 DOI: 10.1038/s41598-017-11472-x

Date:

29-Jun-16 Release date: 02-Aug-17

Protein chain

No UniProt id for this chain

Struc: 474 a.a.

DOI no: 10.1038/s41598-017-11472-x

Sci Rep 7:11217 (2017)

PubMed id: 28894181

Rigidly connected multispecific artificial binders with adjustable geometries.

Y.Wu, A.Batyuk, A.Honegger, F.Brandl, P.R.E.Mittl, A.Plückthun.

ABSTRACT

Multivalent binding proteins can gain biological activities beyond what is inherent in the individual binders, by bringing together different target molecules, restricting their conformational flexibility or changing their subcellular localization. In this study, we demonstrate a method to build up rigid multivalent and multispecific scaffolds by exploiting the modular nature of a repeat protein scaffold and avoiding flexible linkers. We use DARPins (Designed Ankyrin Repeat Proteins), synthetic binding proteins based on the Ankyrin-repeat protein scaffold, as binding units. Their ease of in vitro selection, high production yield and stability make them ideal specificity-conferring building blocks for the design of more complex constructs. C- and N-terminal DARPin capping repeats were re-designed to be joined by a shared helix in such a way that rigid connector modules are formed. This allows us to join two or more DARPins in predefined geometries without compromising their binding affinities and specificities. Nine connector modules with distinct geometries were designed; for eight of these we were able to confirm the structure by X-ray crystallography, while only one did not crystallize. The bispecific constructs were all able to bind both target proteins simultaneously.