title: Crystal structure of the second LNS/LG domain from neurexin 1alpha: Ca2+ binding and the effects of alternative splicing.
abstract: Neurexins mediate protein interactions at the synapse, playing an essential role in synaptic function. Extracellular domains of neurexins, and their fragments, bind a distinct profile of different proteins regulated by alternative splicing and Ca2+. The crystal structure of n1alpha_LNS#2 (the second LNS/LG domain of bovine neurexin 1alpha) reveals large structural differences compared with n1alpha_LNS#6 (or n1beta_LNS), the only other LNS/LG domain for which a structure has been determined. The differences overlap the so-called hyper-variable surface, the putative protein interaction surface that is reshaped as a result of alternative splicing. A Ca2+-binding site is revealed at the center of the hyper-variable surface next to splice insertion sites. Isothermal titration calorimetry indicates that the Ca2+-binding site in n1alpha_LNS#2 has low affinity (Kd approximately 400 microm). Ca2+ binding ceases to be measurable when an 8- or 15-residue splice insert is present at the splice site SS#2 indicating that alternative splicing can affect Ca2+-binding sites of neurexin LNS/LG domains. Our studies initiate a framework for the putative protein interaction sites of neurexin LNS/LG domains. This framework is essential to understand how incorporation of alternative splice inserts expands the information from a limited set of neurexin genes to produce a large array of synaptic adhesion molecules with potentially very different synaptic function.
authors.affiliation: Life Sciences Institute and Department of Pharmacology, The University of Michigan, Ann Arbor, Michigan 48109-2216, USA.
authors.forename: Lauren R
authors.initials: LR
authors.lastname: Sheckler
authors.forename: Lisa
authors.initials: L
authors.lastname: Henry
authors.forename: Shuzo
authors.initials: S
authors.lastname: Sugita
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Südhof
authors.forename: Gabby
authors.initials: G
authors.lastname: Rudenko
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D002118:Calcium / Q000737:chemistry* / Q000378:metabolism; D002417:Cattle; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry*; D007700:Kinetics; D008958:Models, Molecular; D008968:Molecular Conformation; D009479:Neuropeptides / Q000737:chemistry*; D011485:Protein Binding; D017434:Protein Structure, Tertiary
chemical_list: D006023:Glycoproteins; D009479:Neuropeptides; C112162:neurexophilin; D002118:Calcium