Structural basis for cell surface patterning through NetrinG-NGL interactions.

Structural basis for cell surface patterning through NetrinG-NGL interactions.

The EMBO journal (2011-09-29)

Elena Seiradake, Charlotte H Coles, Pavel V Perestenko, Karl Harlos, Robert Andrew Jeffrey McIlhinney, Alexandru Radu Aricescu, Edith Yvonne Jones

PMID21946559

ABSTRACT

Brain wiring depends on cells making highly localized and selective connections through surface protein-protein interactions, including those between NetrinGs and NetrinG ligands (NGLs). The NetrinGs are members of the structurally uncharacterized netrin family. We present a comprehensive crystallographic analysis comprising NetrinG1-NGL1 and NetrinG2-NGL2 complexes, unliganded NetrinG2 and NGL3. Cognate NetrinG-NGL interactions depend on three specificity-conferring NetrinG loops, clasped tightly by matching NGL surfaces. We engineered these NGL surfaces to implant custom-made affinities for NetrinG1 and NetrinG2. In a cellular patterning assay, we demonstrate that NetrinG-binding selectivity can direct the sorting of a mixed population of NGLs into discrete cell surface subdomains. These results provide a molecular model for selectivity-based patterning in a neuronal recognition system, dysregulation of which is associated with severe neuropsychological disorders.

MATERIALS

Product Number

Brand

Product Description

D7777

Sigma-Aldrich

Dulbecco′s Modified Eagle′s Medium - high glucose, With 4500 mg/L glucose, L-glutamine, and sodium pyruvate, without sodium bicarbonate, powder, suitable for cell culture

D5648

Sigma-Aldrich

Dulbecco′s Modified Eagle′s Medium - high glucose, With 4500 mg/L glucose and L-glutamine, without sodium bicarbonate, powder, suitable for cell culture

D1152

Sigma-Aldrich

Dulbecco′s Modified Eagle′s Medium - high glucose, HEPES Modification, With 4500 mg/L glucose, L-glutamine, and 25 mM HEPES, without sodium bicarbonate and pyruvate, powder, suitable for cell culture

D0422

Sigma-Aldrich

Dulbecco′s Modified Eagle′s Medium - high glucose, With 4500 mg/L glucose and sodium bicarbonate, without L-methionine, L-cystine and L-glutamine, liquid, sterile-filtered, suitable for cell culture

D1145

Sigma-Aldrich

Dulbecco′s Modified Eagle′s Medium - high glucose, With 4500 mg/L glucose and sodium bicarbonate, without L-glutamine, sodium pyruvate, and phenol red, liquid, sterile-filtered, suitable for cell culture

D6171

SAFC

Dulbecco′s Modified Eagle′s Medium - high glucose, HEPES modification, With 4500 mg/L glucose, 25 mM HEPES, and sodium bicarbonate, without L-glutamine and sodium pyruvate, liquid, sterile-filtered, suitable for cell culture

D5796

Sigma-Aldrich

Dulbecco′s Modified Eagle′s Medium - high glucose, With 4500 mg/L glucose, L-glutamine, and sodium bicarbonate, without sodium pyruvate, liquid, sterile-filtered, suitable for cell culture

D5671

Sigma-Aldrich

Dulbecco′s Modified Eagle′s Medium - high glucose, With 4500 mg/L glucose and sodium bicarbonate, without L-glutamine and sodium pyruvate, liquid, sterile-filtered, suitable for cell culture, suitable for hybridoma

D0819

Sigma-Aldrich

StableCell^™ DMEM - high glucose, With 4500 mg/L glucose, stable glutamine, and sodium bicarbonate, without sodium pyruvate., liquid, sterile-filtered, suitable for cell culture

D6429

Sigma-Aldrich

Dulbecco′s Modified Eagle′s Medium - high glucose, With 4500 mg/L glucose, L-glutamine, sodium pyruvate, and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture

D6546

Sigma-Aldrich

Dulbecco′s Modified Eagle′s Medium - high glucose, With 4500 mg/L glucose, sodium pyruvate, and sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture

D0822

Sigma-Aldrich

StableCell^™ DMEM - high glucose, With 4500 mg/L glucose, stable glutamine, sodium pyruvate and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture