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Title: Molecular Architecture of Contactin-associated Protein-like 2 (CNTNAP2) and Its Interaction with Contactin 2 (CNTN2)

Abstract

Contactin-associated protein-like 2 (CNTNAP2) is a large multidomain neuronal adhesion molecule implicated in a number of neurological disorders, including epilepsy, schizophrenia, autism spectrum disorder, intellectual disability, and language delay. We reveal in this paper by electron microscopy that the architecture of CNTNAP2 is composed of a large, medium, and small lobe that flex with respect to each other. Using epitope labeling and fragments, we assign the F58C, L1, and L2 domains to the large lobe, the FBG and L3 domains to the middle lobe, and the L4 domain to the small lobe of the CNTNAP2 molecular envelope. Our data reveal that CNTNAP2 has a very different architecture compared with neurexin 1α, a fellow member of the neurexin superfamily and a prototype, suggesting that CNTNAP2 uses a different strategy to integrate into the synaptic protein network. We show that the ectodomains of CNTNAP2 and contactin 2 (CNTN2) bind directly and specifically, with low nanomolar affinity. We show further that mutations in CNTNAP2 implicated in autism spectrum disorder are not segregated but are distributed over the whole ectodomain. Finally, the molecular shape and dimensions of CNTNAP2 place constraints on how CNTNAP2 integrates in the cleft of axo-glial and neuronal contact sites andmore » how it functions as an organizing and adhesive molecule.« less

Authors:
 [1];  [2];  [3];  [2];  [4];  [3];  [5];  [5];  [6];  [7];  [2];  [2];  [3];  [2]
  1. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Molecular Foundry; Xi'an Jiaotong Univ., Xi'an (China). Frontier Inst. of Science and Technology. School of Life Science and Technology. Key Lab. of Biomedical Information Engineering of Ministry of Education. Center for Mitochondrial Biology and Medicine
  2. Univ. of Texas Medical Branch, Galveston, TX (United States). Dept. of Pharmacology and Toxicology. Sealy Center for Structural Biology and Molecular Biophysics
  3. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Molecular Foundry
  4. Xi'an Jiaotong Univ., Xi'an (China). Frontier Inst. of Science and Technology. School of Life Science and Technology. Key Lab. of Biomedical Information Engineering of Ministry of Education. Center for Mitochondrial Biology and Medicine
  5. Univ. of Michigan, Ann Arbor, MI (United States)
  6. Univ. of Texas Medical Branch, Galveston, TX (United States). Sealy Center for Structural Biology and Molecular Biophysics
  7. Univ. of Texas Medical Branch, Galveston, TX (United States). Sealy Center for Structural Biology and Molecular Biophysics. Dept. of Biochemistry and Molecular Biology
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of Texas Medical Branch, Galveston, TX (United States); Xi'an Jiaotong Univ., Xi'an (China)
Sponsoring Org.:
USDOE; National Inst. of Health (NIH) (United States); Univ. of Texas (United States); Brain and Behavior Research Foundation (United States); Ministry of Science and Technology (China)
OSTI Identifier:
1377581
Grant/Contract Number:
AC02-05CH11231; R01MH077303; 2015CB553602
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Journal of Biological Chemistry
Additional Journal Information:
Journal Volume: 291; Journal Issue: 46; Journal ID: ISSN 0021-9258
Publisher:
American Society for Biochemistry and Molecular Biology
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; cell adhesion; cell surface receptor; protein-protein interaction; structural biology synapse; contactin; contactin-associated protein like; neuropsychiatric disorders; single particle analysis; synaptic organizer

Citation Formats

Lu, Zhuoyang, Reddy, M. V. V. V. Sekhar, Liu, Jianfang, Kalichava, Ana, Liu, Jiankang, Zhang, Lei, Chen, Fang, Wang, Yun, Holthauzen, Luis Marcelo F., White, Mark A., Seshadrinathan, Suchithra, Zhong, Xiaoying, Ren, Gang, and Rudenko, Gabby. Molecular Architecture of Contactin-associated Protein-like 2 (CNTNAP2) and Its Interaction with Contactin 2 (CNTN2). United States: N. p., 2016. Web. doi:10.1074/jbc.M116.748236.
Lu, Zhuoyang, Reddy, M. V. V. V. Sekhar, Liu, Jianfang, Kalichava, Ana, Liu, Jiankang, Zhang, Lei, Chen, Fang, Wang, Yun, Holthauzen, Luis Marcelo F., White, Mark A., Seshadrinathan, Suchithra, Zhong, Xiaoying, Ren, Gang, & Rudenko, Gabby. Molecular Architecture of Contactin-associated Protein-like 2 (CNTNAP2) and Its Interaction with Contactin 2 (CNTN2). United States. doi:10.1074/jbc.M116.748236.
Lu, Zhuoyang, Reddy, M. V. V. V. Sekhar, Liu, Jianfang, Kalichava, Ana, Liu, Jiankang, Zhang, Lei, Chen, Fang, Wang, Yun, Holthauzen, Luis Marcelo F., White, Mark A., Seshadrinathan, Suchithra, Zhong, Xiaoying, Ren, Gang, and Rudenko, Gabby. 2016. "Molecular Architecture of Contactin-associated Protein-like 2 (CNTNAP2) and Its Interaction with Contactin 2 (CNTN2)". United States. doi:10.1074/jbc.M116.748236. https://www.osti.gov/servlets/purl/1377581.
@article{osti_1377581,
title = {Molecular Architecture of Contactin-associated Protein-like 2 (CNTNAP2) and Its Interaction with Contactin 2 (CNTN2)},
author = {Lu, Zhuoyang and Reddy, M. V. V. V. Sekhar and Liu, Jianfang and Kalichava, Ana and Liu, Jiankang and Zhang, Lei and Chen, Fang and Wang, Yun and Holthauzen, Luis Marcelo F. and White, Mark A. and Seshadrinathan, Suchithra and Zhong, Xiaoying and Ren, Gang and Rudenko, Gabby},
abstractNote = {Contactin-associated protein-like 2 (CNTNAP2) is a large multidomain neuronal adhesion molecule implicated in a number of neurological disorders, including epilepsy, schizophrenia, autism spectrum disorder, intellectual disability, and language delay. We reveal in this paper by electron microscopy that the architecture of CNTNAP2 is composed of a large, medium, and small lobe that flex with respect to each other. Using epitope labeling and fragments, we assign the F58C, L1, and L2 domains to the large lobe, the FBG and L3 domains to the middle lobe, and the L4 domain to the small lobe of the CNTNAP2 molecular envelope. Our data reveal that CNTNAP2 has a very different architecture compared with neurexin 1α, a fellow member of the neurexin superfamily and a prototype, suggesting that CNTNAP2 uses a different strategy to integrate into the synaptic protein network. We show that the ectodomains of CNTNAP2 and contactin 2 (CNTN2) bind directly and specifically, with low nanomolar affinity. We show further that mutations in CNTNAP2 implicated in autism spectrum disorder are not segregated but are distributed over the whole ectodomain. Finally, the molecular shape and dimensions of CNTNAP2 place constraints on how CNTNAP2 integrates in the cleft of axo-glial and neuronal contact sites and how it functions as an organizing and adhesive molecule.},
doi = {10.1074/jbc.M116.748236},
journal = {Journal of Biological Chemistry},
number = 46,
volume = 291,
place = {United States},
year = 2016,
month = 9
}

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