Structure-function analysis of Nel, a thrombospondin-1-like glycoprotein involved in neural development and functions

Ritsuko Nakamura, Chizu Nakamoto, Hiroya Obama, Elaine Durward, Masaru Nakamoto

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Nel (neural epidermal growth factor (EGF)-like molecule) is a multimeric, multimodular extracellular glycoprotein with heparin-binding activity and structural similarities to thrombospondin-1. Nel is predominantly expressed in the nervous system and has been implicated in neuronal proliferation and differentiation, retinal axon guidance, synaptic functions, and spatial learning. The Nel protein contains an N-terminal thrombospondin-1 (TSP-N) domain, five cysteine-rich domains, and six EGF-like domains. However, little is known about the functions of specific domains of the Nel protein. In this study, we have performed structure-function analysis of Nel, by using a series of expression constructs for different regions of the Nel protein. Our studies demonstrate that the TSP-N domain is responsible for homo-multimer formation of Nel and its heparin-binding activity. In vivo, Nel and related Nell1 are expressed in several regions of the mouse central nervous system with partly overlapping patterns. When they are expressed in the same cells in vitro, Nel and Nell1 can form hetero-multimers through the TSP-N domain, but they do not hetero-oligomerize with thrombospondin-1. Whereas both the TSP-N domain and cysteine-rich domains can bind to retinal axons in vivo, only the latter causes growth cone collapse in cultured retinal axons, suggesting that cysteine-rich domains interact with and activate an inhibitory axon guidance receptor. These results suggest that Nel interacts with a range of molecules through its different domains and exerts distinct functions.

Original languageEnglish
Pages (from-to)3282-3291
Number of pages10
JournalThe Journal of Biological Chemistry
Volume287
Issue number5
Early online date8 Dec 2011
DOIs
Publication statusPublished - 27 Jan 2012

Fingerprint

Thrombospondin 1
Glycoproteins
Cysteine
Neurology
Epidermal Growth Factor
Axons
Heparin
Growth Cones
Proteins
Molecules
Nervous System
Cones
Central Nervous System

Keywords

  • axon
  • extracellular matrix proteins
  • heparin binding protein
  • neurite outgrowth
  • protein domains
  • protein-protein interactions
  • Nel

Cite this

Structure-function analysis of Nel, a thrombospondin-1-like glycoprotein involved in neural development and functions. / Nakamura, Ritsuko; Nakamoto, Chizu; Obama, Hiroya; Durward, Elaine; Nakamoto, Masaru.

In: The Journal of Biological Chemistry, Vol. 287, No. 5, 27.01.2012, p. 3282-3291.

Research output: Contribution to journalArticle

Nakamura, Ritsuko ; Nakamoto, Chizu ; Obama, Hiroya ; Durward, Elaine ; Nakamoto, Masaru. / Structure-function analysis of Nel, a thrombospondin-1-like glycoprotein involved in neural development and functions. In: The Journal of Biological Chemistry. 2012 ; Vol. 287, No. 5. pp. 3282-3291.
@article{a46eb70e28af4d35999b52e8fe8328de,
title = "Structure-function analysis of Nel, a thrombospondin-1-like glycoprotein involved in neural development and functions",
abstract = "Nel (neural epidermal growth factor (EGF)-like molecule) is a multimeric, multimodular extracellular glycoprotein with heparin-binding activity and structural similarities to thrombospondin-1. Nel is predominantly expressed in the nervous system and has been implicated in neuronal proliferation and differentiation, retinal axon guidance, synaptic functions, and spatial learning. The Nel protein contains an N-terminal thrombospondin-1 (TSP-N) domain, five cysteine-rich domains, and six EGF-like domains. However, little is known about the functions of specific domains of the Nel protein. In this study, we have performed structure-function analysis of Nel, by using a series of expression constructs for different regions of the Nel protein. Our studies demonstrate that the TSP-N domain is responsible for homo-multimer formation of Nel and its heparin-binding activity. In vivo, Nel and related Nell1 are expressed in several regions of the mouse central nervous system with partly overlapping patterns. When they are expressed in the same cells in vitro, Nel and Nell1 can form hetero-multimers through the TSP-N domain, but they do not hetero-oligomerize with thrombospondin-1. Whereas both the TSP-N domain and cysteine-rich domains can bind to retinal axons in vivo, only the latter causes growth cone collapse in cultured retinal axons, suggesting that cysteine-rich domains interact with and activate an inhibitory axon guidance receptor. These results suggest that Nel interacts with a range of molecules through its different domains and exerts distinct functions.",
keywords = "axon, extracellular matrix proteins, heparin binding protein, neurite outgrowth, protein domains, protein-protein interactions, Nel",
author = "Ritsuko Nakamura and Chizu Nakamoto and Hiroya Obama and Elaine Durward and Masaru Nakamoto",
year = "2012",
month = "1",
day = "27",
doi = "10.1074/jbc.M111.281485",
language = "English",
volume = "287",
pages = "3282--3291",
journal = "The Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC",
number = "5",

}

TY - JOUR

T1 - Structure-function analysis of Nel, a thrombospondin-1-like glycoprotein involved in neural development and functions

AU - Nakamura, Ritsuko

AU - Nakamoto, Chizu

AU - Obama, Hiroya

AU - Durward, Elaine

AU - Nakamoto, Masaru

PY - 2012/1/27

Y1 - 2012/1/27

N2 - Nel (neural epidermal growth factor (EGF)-like molecule) is a multimeric, multimodular extracellular glycoprotein with heparin-binding activity and structural similarities to thrombospondin-1. Nel is predominantly expressed in the nervous system and has been implicated in neuronal proliferation and differentiation, retinal axon guidance, synaptic functions, and spatial learning. The Nel protein contains an N-terminal thrombospondin-1 (TSP-N) domain, five cysteine-rich domains, and six EGF-like domains. However, little is known about the functions of specific domains of the Nel protein. In this study, we have performed structure-function analysis of Nel, by using a series of expression constructs for different regions of the Nel protein. Our studies demonstrate that the TSP-N domain is responsible for homo-multimer formation of Nel and its heparin-binding activity. In vivo, Nel and related Nell1 are expressed in several regions of the mouse central nervous system with partly overlapping patterns. When they are expressed in the same cells in vitro, Nel and Nell1 can form hetero-multimers through the TSP-N domain, but they do not hetero-oligomerize with thrombospondin-1. Whereas both the TSP-N domain and cysteine-rich domains can bind to retinal axons in vivo, only the latter causes growth cone collapse in cultured retinal axons, suggesting that cysteine-rich domains interact with and activate an inhibitory axon guidance receptor. These results suggest that Nel interacts with a range of molecules through its different domains and exerts distinct functions.

AB - Nel (neural epidermal growth factor (EGF)-like molecule) is a multimeric, multimodular extracellular glycoprotein with heparin-binding activity and structural similarities to thrombospondin-1. Nel is predominantly expressed in the nervous system and has been implicated in neuronal proliferation and differentiation, retinal axon guidance, synaptic functions, and spatial learning. The Nel protein contains an N-terminal thrombospondin-1 (TSP-N) domain, five cysteine-rich domains, and six EGF-like domains. However, little is known about the functions of specific domains of the Nel protein. In this study, we have performed structure-function analysis of Nel, by using a series of expression constructs for different regions of the Nel protein. Our studies demonstrate that the TSP-N domain is responsible for homo-multimer formation of Nel and its heparin-binding activity. In vivo, Nel and related Nell1 are expressed in several regions of the mouse central nervous system with partly overlapping patterns. When they are expressed in the same cells in vitro, Nel and Nell1 can form hetero-multimers through the TSP-N domain, but they do not hetero-oligomerize with thrombospondin-1. Whereas both the TSP-N domain and cysteine-rich domains can bind to retinal axons in vivo, only the latter causes growth cone collapse in cultured retinal axons, suggesting that cysteine-rich domains interact with and activate an inhibitory axon guidance receptor. These results suggest that Nel interacts with a range of molecules through its different domains and exerts distinct functions.

KW - axon

KW - extracellular matrix proteins

KW - heparin binding protein

KW - neurite outgrowth

KW - protein domains

KW - protein-protein interactions

KW - Nel

U2 - 10.1074/jbc.M111.281485

DO - 10.1074/jbc.M111.281485

M3 - Article

VL - 287

SP - 3282

EP - 3291

JO - The Journal of Biological Chemistry

JF - The Journal of Biological Chemistry

SN - 0021-9258

IS - 5

ER -