Characterization of the receptors for axon guidance factor netrin-4 and identification of the binding domains.

Characterization of the receptors for axon guidance factor netrin-4 and identification of the binding domains. Research Abstract Details 

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Characterization of the receptors for axon guidance factor netrin-4 and identification of the binding domains. Abstract Text:

Shutong Qin,Lihong Yu,Yan Gao,Renping Zhou,Chenggang Zhang,

Netrins are a family of secreted protein related to laminin and act as tropic cues directing axon growth and cell migration during neural development. Netrin-4 is a novel member of netrin family recently identified in the vertebrate with neuritis elongation promoting activity; however, the receptors for netrin-4 are still unknown. To better understand the function and signal transduction pathway of netrin-4, the potential receptors for netrin-4 were studied in this paper. The netrin-4 protein was prepared by introducing a eukaryotic expression vector with a secretable alkaline phosphatase tag (AP4) into COS7 cells to allow the expression of AP4-netrin4 fusion protein. Axon guidance activity of netrin-4 was confirmed by using the cortical explants. After incubation with cultured primary cortical neurons, the neurons were distinctly labeled by the AP4-coupled netrin-4 ligands. In contrast, the binding activity of AP4-netrin4 to neurons could be completely competed by the exogenously expressed netrin-4 protein without AP4 tag, indicating specificity of netrin-4 binding to the potential receptors. Moreover, netrin-4 could also bind to CHO cells transfected with the plasmids expressing two known receptors for netrin-1, Deleted in Colorectal Cancer (DCC) and UNC5 homolog 1 (UNC5H1) respectively. As there are three domains in netrin-4, we further tried to narrow down the region containing binding sites with the receptors. Interestingly, only the N-terminal domain (LNT) could bind to DCC and UNC5H1. A further ligand-receptor binding analysis showed that both the N- and the C-terminal domain (NCT) but not the EGF-like (EGFL) domain of netrin-4 could bind to the surface of cultured primary neurons, indicating the existence of novel receptors for netrin-4. After competed by netrin-4, we confirmed that the binding of AP tagged netrin-4 domains to neurons were also netrin-4 dependent. The binding activity of the N-terminal domain of netrin-4 is about 3-fold higher than that for the C-terminal domain. In summary, our data here indicated that the two known receptors for netrin-1, DCC and UNC5H1, are also receptors for netrin-4, while only LNT but not EGFL and NCT is the key domain for specific binding. In addition, there are novel receptors for netrin-4, where both LNT and NCT but not EGFL are key domains for binding.

Characterization of the receptors for axon guidance factor netrin-4 and identification of the binding domains. Publishing Authors By Initials

S Qin,L Yu,Y Gao,R Zhou,C Zhang,

For similar proteins: recombinant proteins: recombinant fusion proteins research abstracts see: proteins: recombinant proteins: recombinant fusion proteins research

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MEDLINE DATE:

Characterization of the receptors for axon guidance factor netrin-4 and identification of the binding domains. Journal Published:

PUBLICATION TYPE: Research Support, U.S. Gov't,

Journal: Molecular and cellular neurosciences

VOLUME: 34

Page Numbers: 243-50

Journal Abbreviation: Mol. Cell. Neurosci.

ISSN: 1044-7431

DAY: 15

MONTH: 12

YEAR: 2006

Characterization of the receptors for axon guidance factor netrin-4 and identification of the binding domains. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 9100095

Characterization of the receptors for axon guidance factor netrin-4 and identification of the binding domains. Keywords Mesh Terms:

KEYWORDS: Recombinant Fusion Proteins

MESH TERMS: metabolism

Chemical & Substance for Abstract: Characterization of the receptors for axon guidance factor netrin-4 and identification of the binding domains. Information

Substance Name: Alkaline Phosphatase

Registry Number: EC 3.1.3.1

Grant and Affiliation Information for Characterization of the receptors for axon guidance factor netrin-4 and identification of the binding domains.

AFFILIATION: Department of Neurobiology, Beijing Institute of Radiation Medicine, Taiping Road 27, Beijing 100850, China.

Country: United States

AGENCY: United States NICHD

GRANT: P01 HD23315

ACRONYM: HD

MEDLINETA: Mol Cell Neurosci

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