Calsyntenin-3 molecular architecture and interaction with neurexin 1α

Zhuoyang Lu, Yun Wang, Fang Chen, Huimin Tong, M. V V V Sekhar Reddy, Lin Luo, Suchithra Seshadrinathan, Lei Zhang, Luis Marcelo F Holthauzen, Ann Marie Craig, Gang Ren, Gabrielle Rudenko

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Calsyntenin 3 (Cstn3 or Clstn3), a recently identified synaptic organizer, promotes the development of synapses. Cstn3 localizes to the postsynaptic membrane and triggers presynaptic differentiation. Calsyntenin members play an evolutionarily conserved role in memory and learning. Cstn3 was recently shown in cell-based assays to interact with neurexin 1α (n1α), a synaptic organizer that is implicated in neuropsychiatric disease. Interaction would permit Cstn3 and n1α to form a trans-synaptic complex and promote synaptic differentiation. However, it is contentious whether Cstn3 binds n1α directly. To understand the structure and function of Cstn3, we determined its architecture by electron microscopy and delineated the interaction between Cstn3 and n1α biochemically and biophysically. We show that Cstn3 ectodomains form monomers as well as tetramers that are stabilized by disulfide bonds and Ca2+, and both are probably flexible in solution. We show further that the extracellular domains of Cstn3 and n1α interact directly and that both Cstn3 monomers and tetramers bind n1α with nanomolar affinity. The interaction is promoted by Ca2+ and requires minimally the LNS domain of Cstn3. Furthermore, Cstn3 uses a fundamentally different mechanism to bind n1α compared with other neurexin partners, such as the synaptic organizer neuroligin 2, because Cstn3 does not strictly require the sixth LNS domain of n1α. Our structural data suggest how Cstn3 as a synaptic organizer on the postsynaptic membrane, particularly in tetrameric form, may assemble radially symmetric trans-synaptic bridges with the presynaptic synaptic organizer n1α to recruit and spatially organize proteins into networks essential for synaptic function.

Original languageEnglish (US)
Pages (from-to)34530-34542
Number of pages13
JournalJournal of Biological Chemistry
Volume289
Issue number50
DOIs
StatePublished - Dec 12 2014

Fingerprint

Monomers
Membranes
Disulfides
Synapses
Electron microscopy
Assays
Electron Microscopy
Learning
Data storage equipment
Proteins
neuroligin 2

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Calsyntenin-3 molecular architecture and interaction with neurexin 1α. / Lu, Zhuoyang; Wang, Yun; Chen, Fang; Tong, Huimin; Reddy, M. V V V Sekhar; Luo, Lin; Seshadrinathan, Suchithra; Zhang, Lei; Holthauzen, Luis Marcelo F; Craig, Ann Marie; Ren, Gang; Rudenko, Gabrielle.

In: Journal of Biological Chemistry, Vol. 289, No. 50, 12.12.2014, p. 34530-34542.

Research output: Contribution to journalArticle

Lu, Z, Wang, Y, Chen, F, Tong, H, Reddy, MVVVS, Luo, L, Seshadrinathan, S, Zhang, L, Holthauzen, LMF, Craig, AM, Ren, G & Rudenko, G 2014, 'Calsyntenin-3 molecular architecture and interaction with neurexin 1α', Journal of Biological Chemistry, vol. 289, no. 50, pp. 34530-34542. https://doi.org/10.1074/jbc.M114.606806
Lu, Zhuoyang ; Wang, Yun ; Chen, Fang ; Tong, Huimin ; Reddy, M. V V V Sekhar ; Luo, Lin ; Seshadrinathan, Suchithra ; Zhang, Lei ; Holthauzen, Luis Marcelo F ; Craig, Ann Marie ; Ren, Gang ; Rudenko, Gabrielle. / Calsyntenin-3 molecular architecture and interaction with neurexin 1α. In: Journal of Biological Chemistry. 2014 ; Vol. 289, No. 50. pp. 34530-34542.
@article{23f8bd81cf44405e92efb96ef9683d4a,
title = "Calsyntenin-3 molecular architecture and interaction with neurexin 1α",
abstract = "Calsyntenin 3 (Cstn3 or Clstn3), a recently identified synaptic organizer, promotes the development of synapses. Cstn3 localizes to the postsynaptic membrane and triggers presynaptic differentiation. Calsyntenin members play an evolutionarily conserved role in memory and learning. Cstn3 was recently shown in cell-based assays to interact with neurexin 1α (n1α), a synaptic organizer that is implicated in neuropsychiatric disease. Interaction would permit Cstn3 and n1α to form a trans-synaptic complex and promote synaptic differentiation. However, it is contentious whether Cstn3 binds n1α directly. To understand the structure and function of Cstn3, we determined its architecture by electron microscopy and delineated the interaction between Cstn3 and n1α biochemically and biophysically. We show that Cstn3 ectodomains form monomers as well as tetramers that are stabilized by disulfide bonds and Ca2+, and both are probably flexible in solution. We show further that the extracellular domains of Cstn3 and n1α interact directly and that both Cstn3 monomers and tetramers bind n1α with nanomolar affinity. The interaction is promoted by Ca2+ and requires minimally the LNS domain of Cstn3. Furthermore, Cstn3 uses a fundamentally different mechanism to bind n1α compared with other neurexin partners, such as the synaptic organizer neuroligin 2, because Cstn3 does not strictly require the sixth LNS domain of n1α. Our structural data suggest how Cstn3 as a synaptic organizer on the postsynaptic membrane, particularly in tetrameric form, may assemble radially symmetric trans-synaptic bridges with the presynaptic synaptic organizer n1α to recruit and spatially organize proteins into networks essential for synaptic function.",
author = "Zhuoyang Lu and Yun Wang and Fang Chen and Huimin Tong and Reddy, {M. V V V Sekhar} and Lin Luo and Suchithra Seshadrinathan and Lei Zhang and Holthauzen, {Luis Marcelo F} and Craig, {Ann Marie} and Gang Ren and Gabrielle Rudenko",
year = "2014",
month = "12",
day = "12",
doi = "10.1074/jbc.M114.606806",
language = "English (US)",
volume = "289",
pages = "34530--34542",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "50",

}

TY - JOUR

T1 - Calsyntenin-3 molecular architecture and interaction with neurexin 1α

AU - Lu, Zhuoyang

AU - Wang, Yun

AU - Chen, Fang

AU - Tong, Huimin

AU - Reddy, M. V V V Sekhar

AU - Luo, Lin

AU - Seshadrinathan, Suchithra

AU - Zhang, Lei

AU - Holthauzen, Luis Marcelo F

AU - Craig, Ann Marie

AU - Ren, Gang

AU - Rudenko, Gabrielle

PY - 2014/12/12

Y1 - 2014/12/12

N2 - Calsyntenin 3 (Cstn3 or Clstn3), a recently identified synaptic organizer, promotes the development of synapses. Cstn3 localizes to the postsynaptic membrane and triggers presynaptic differentiation. Calsyntenin members play an evolutionarily conserved role in memory and learning. Cstn3 was recently shown in cell-based assays to interact with neurexin 1α (n1α), a synaptic organizer that is implicated in neuropsychiatric disease. Interaction would permit Cstn3 and n1α to form a trans-synaptic complex and promote synaptic differentiation. However, it is contentious whether Cstn3 binds n1α directly. To understand the structure and function of Cstn3, we determined its architecture by electron microscopy and delineated the interaction between Cstn3 and n1α biochemically and biophysically. We show that Cstn3 ectodomains form monomers as well as tetramers that are stabilized by disulfide bonds and Ca2+, and both are probably flexible in solution. We show further that the extracellular domains of Cstn3 and n1α interact directly and that both Cstn3 monomers and tetramers bind n1α with nanomolar affinity. The interaction is promoted by Ca2+ and requires minimally the LNS domain of Cstn3. Furthermore, Cstn3 uses a fundamentally different mechanism to bind n1α compared with other neurexin partners, such as the synaptic organizer neuroligin 2, because Cstn3 does not strictly require the sixth LNS domain of n1α. Our structural data suggest how Cstn3 as a synaptic organizer on the postsynaptic membrane, particularly in tetrameric form, may assemble radially symmetric trans-synaptic bridges with the presynaptic synaptic organizer n1α to recruit and spatially organize proteins into networks essential for synaptic function.

AB - Calsyntenin 3 (Cstn3 or Clstn3), a recently identified synaptic organizer, promotes the development of synapses. Cstn3 localizes to the postsynaptic membrane and triggers presynaptic differentiation. Calsyntenin members play an evolutionarily conserved role in memory and learning. Cstn3 was recently shown in cell-based assays to interact with neurexin 1α (n1α), a synaptic organizer that is implicated in neuropsychiatric disease. Interaction would permit Cstn3 and n1α to form a trans-synaptic complex and promote synaptic differentiation. However, it is contentious whether Cstn3 binds n1α directly. To understand the structure and function of Cstn3, we determined its architecture by electron microscopy and delineated the interaction between Cstn3 and n1α biochemically and biophysically. We show that Cstn3 ectodomains form monomers as well as tetramers that are stabilized by disulfide bonds and Ca2+, and both are probably flexible in solution. We show further that the extracellular domains of Cstn3 and n1α interact directly and that both Cstn3 monomers and tetramers bind n1α with nanomolar affinity. The interaction is promoted by Ca2+ and requires minimally the LNS domain of Cstn3. Furthermore, Cstn3 uses a fundamentally different mechanism to bind n1α compared with other neurexin partners, such as the synaptic organizer neuroligin 2, because Cstn3 does not strictly require the sixth LNS domain of n1α. Our structural data suggest how Cstn3 as a synaptic organizer on the postsynaptic membrane, particularly in tetrameric form, may assemble radially symmetric trans-synaptic bridges with the presynaptic synaptic organizer n1α to recruit and spatially organize proteins into networks essential for synaptic function.

UR - http://www.scopus.com/inward/record.url?scp=84919338468&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84919338468&partnerID=8YFLogxK

U2 - 10.1074/jbc.M114.606806

DO - 10.1074/jbc.M114.606806

M3 - Article

VL - 289

SP - 34530

EP - 34542

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 50

ER -