Crystal structure of the extracellular cholinesterase-like domain from neuroligin-2

Koehnke, Jesko; Jin, Xiangshu; Budreck, Elaine C.; Posy, Shoshana; Scheiffele, Peter; Shapiro, Lawrence

doi:10.1073/pnas.0711701105

Cited by 38 publications

(36 citation statements)

References 36 publications

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“…First, we probed the degenerate EF hand (30) for its contribution to complex formation, but deletion (Nlgn1ϩB⌬EF1) did not alter binding to ␤LNS (Fig. S10), thereby validating recent Nlgn crystal structures that did not detect Ca 2ϩ bound to this motif (15)(16)(17)31). Next, we selected four presumptive interface sites in Nlgn that match the criteria analyzed above, i.e., (i) the ϩSS4 and ϩB inserts may interfere to reduce (but not abolish) binding, (ii) the contact site is likely hydrophobic, and (iii) it should contain a residue capable of joining the Ca 2ϩ coordination of Nrxn.…”

Section: Resultsmentioning

confidence: 99%

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Mutational analysis of the neurexin/neuroligin complex reveals essential and regulatory components

Reißner

Klose

Fairless

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

103

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Neurexins are cell-surface molecules that bind neuroligins to form a heterophilic, Ca 2؉ -dependent complex at central synapses. This transsynaptic complex is required for efficient neurotransmission and is involved in the formation of synaptic contacts. In addition, both molecules have been identified as candidate genes for autism. Here we performed mutagenesis experiments to probe for essential components of the neurexin/neuroligin binding interface at the single-amino acid level. We found that in neurexins the contact area is sharply delineated and consists of hydrophobic residues of the LNS domain that surround a Ca 2؉ binding pocket. Point mutations that changed electrostatic and shape properties leave Ca 2؉ coordination intact but completely inhibit neuroligin binding, whereas alternative splicing in ␣-and ␤-neurexins and in neuroligins has a weaker effect on complex formation. In neuroligins, the contact area appears less distinct because exchange of a more distant aspartate completely abolished binding to neurexin but many mutations of predicted interface residues had no strong effect on binding. Together with calculations of energy terms for presumed interface hot spots that complement and extend our mutagenesis and recent crystal structure data, this study presents a comprehensive structural basis for the complex formation of neurexins and neuroligins and their transsynaptic signaling between neurons.calcium ͉ cell adhesion ͉ LNS domain ͉ neurotransmission ͉ synaptogenesis T he heterophilic complex formed by cell adhesion molecules neurexin (Nrxn) and neuroligin (Nlgn) reflects the asymmetric nature of the synapse with presynaptic and postsynaptic specializations (1, 2). Nrxns and Nlgns are essential molecules because they perform important functions in synaptic transmission (3, 4) and differentiation of synaptic contacts (5, 6), and both molecules have been identified as candidate genes for autism (7,8).Nrxns form a family of transmembrane proteins with variable extracellular sequences. Nrxn genes (Nrxn1-3) give rise to ␣-neurexins and shorter ␤-neurexins that contain five (␣-Nrxn) or two (␤-Nrxn) splice sites (SS1-5) (9). Although they share most sequences, the essential role of ␣-Nrxn in neurotransmission cannot be replaced by ␤-Nrxn (10), and one ligand exists for ␣-Nrxn that does not bind to ␤-Nrxn (11). In contrast, Nlgn was discovered by its interaction with ␤-Nrxn (12). The cholinesterase-like adhesion molecule (CAM) domain of Nlgn interacts with the extracellular domain of ␤-Nrxn in a Ca 2ϩ -dependent manner, and binding is facilitated by the splice variation of ␤-Nrxn that lacks an insert in SS4 (13). Nlgn mRNA is also susceptible to splicing, at two positions referred to as A and B (12), including splice variants with no insert in B that bind to all ␤-Nrxns and presumably ␣-Nrxn (14). Therefore, the Nrxn/Nlgn complex involves a domain shared by ␣-and ␤-Nrxns, and any structural characterization needs to account for the Ca 2ϩ dependence and regulation by alternative splicing (15-17).Extracell...

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Section: Resultsmentioning

confidence: 99%

“…Because of available data from cocrystal structures of the Nrxn/Nlgn complex (15)(16)(17)31), the contribution of suspected residues to complex stability can be calculated and compared with mutagenesis analyses (Fig. 6).…”

Section: Resultsmentioning

confidence: 99%

Mutational analysis of the neurexin/neuroligin complex reveals essential and regulatory components

Reißner

Klose

Fairless

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

103

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“…The neuroligin dimer was initially inferred from similarity to the protein sequence of acetylcholinesterase [28], and the presence of a dimer has been supported both biochemically as well as functionally: mutations in the extracellular domain based on the acetylcholinesterase structure abolish both the biochemical dimer and the overexpression phenotype of the molecule [29]. When the crystal structure of the neuroligin extracellular domain was later determined, it confirmed dimerization highly reminiscent of cholinesterases [30][31][32][33]. With structure in hand, function was revisited: a series of mutations that abolish dimerization were found to induce a corresponding reduction in function [34].…”

Section: Dimerizationmentioning

confidence: 94%

The cellular and molecular landscape of neuroligins

Bemben

Shipman

Nicoll

et al. 2015

Trends in Neurosciences

153

137

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“…3B) and we compared their different exonic structures to the secondary structure elements of crystallized Neuroligins (Fabrichny et al, 2007;Koehnke et al, 2008a). In Neuroligins, most of the intron gain/loss events are localized outside or at the borders of secondary structures and only few events are found inside alphahelices.…”

Section: Correlation Of Intron Gain/ Loss Events and Secondary Proteimentioning

confidence: 99%

Characterization of the neuroligin gene family expression and evolution in zebrafish

Rissone¹,

Sangiorgio²,

Monopoli³

et al. 2010

Developmental Dynamics

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Neuroligins constitute a family of transmembrane proteins localized at the postsynaptic side of both excitatory and inhibitory synapses of the central nervous system. They are involved in synaptic function and maturation and recent studies have linked mutations in specific human Neuroligins to mental retardation and autism. We isolated the human Neuroligin homologs in Danio rerio. Next, we studied their gene structures and we reconstructed the evolution of the Neuroligin genes across vertebrate phyla. Using reverse-transcriptase polymerase chain reaction, we analyzed the expression and alternative splicing pattern of each gene during zebrafish embryonic development and in different adult organs. By in situ hybridization, we analyzed the temporal and spatial expression pattern during embryonic development and larval stages and we found that zebrafish Neuroligins are expressed throughout the nervous system. Globally, our results indicate that, during evolution, specific subfunctionalization events occurred within paralogous members of this gene family in zebrafish. Developmental Dynamics 239:688-702,

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Crystal structure of the extracellular cholinesterase-like domain from neuroligin-2

Cited by 38 publications

References 36 publications

Mutational analysis of the neurexin/neuroligin complex reveals essential and regulatory components

Mutational analysis of the neurexin/neuroligin complex reveals essential and regulatory components

The cellular and molecular landscape of neuroligins

Characterization of the neuroligin gene family expression and evolution in zebrafish

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