Design and structural characterisation of olfactomedin-1 variants as tools for functional studies

Abstract Background Olfactomedin-1 (Olfm1; also known as Noelin or Pancortin) is a highly-expressed secreted brain and retina protein and its four isoforms have different roles in nervous system development and function. Structural studies showed that the long Olfm1 isoform BMZ forms a disulfide-lin...

Full description

Bibliographic Details
Main Authors: Matti F. Pronker, Hugo van den Hoek, Bert J. C. Janssen
Format: Article
Language:English
Published: BMC 2019-11-01
Series:BMC Molecular and Cell Biology
Subjects:
Online Access:http://link.springer.com/article/10.1186/s12860-019-0232-1
id doaj-e4c38507be8b460d94ef7e5fbfbe5896
record_format Article
spelling doaj-e4c38507be8b460d94ef7e5fbfbe58962020-11-25T04:02:18ZengBMCBMC Molecular and Cell Biology2661-88502019-11-0120111510.1186/s12860-019-0232-1Design and structural characterisation of olfactomedin-1 variants as tools for functional studiesMatti F. Pronker0Hugo van den Hoek1Bert J. C. Janssen2MRC Laboratory of Molecular Biology, Division of NeurobiologyBijvoet Center for Biomolecular Research, Utrecht University, Crystal and Structural Chemistry, KruytgebouwBijvoet Center for Biomolecular Research, Utrecht University, Crystal and Structural Chemistry, KruytgebouwAbstract Background Olfactomedin-1 (Olfm1; also known as Noelin or Pancortin) is a highly-expressed secreted brain and retina protein and its four isoforms have different roles in nervous system development and function. Structural studies showed that the long Olfm1 isoform BMZ forms a disulfide-linked tetramer with a V-shaped architecture. The tips of the Olfm1 “V” each consist of two C-terminal β-propeller domains that enclose a calcium binding site. Functional characterisation of Olfm1 may be aided by new biochemical tools derived from these core structural elements. Results Here we present the production, purification and structural analysis of three novel monomeric, dimeric and tetrameric forms of mammalian Olfm1 for functional studies. We characterise these constructs structurally by high-resolution X-ray crystallography and small-angle X-ray scattering. The crystal structure of the Olfm1 β-propeller domain (to 1.25 Å) represents the highest-resolution structure of an olfactomedin family member to date, revealing features such as a hydrophilic tunnel containing water molecules running into the core of the domain where the calcium binding site resides. The shorter Olfactomedin-1 isoform BMY is a disulfide-linked tetramer with a shape similar to the corresponding region in the longer BMZ isoform. Conclusions These recombinantly-expressed protein tools should assist future studies, for example of biophysical, electrophysiological or morphological nature, to help elucidate the functions of Olfm1 in the mature mammalian brain. The control over the oligomeric state of Olfm1 provides a firm basis to better understand the role of Olfm1 in the (trans-synaptic) tethering or avidity-mediated clustering of synaptic receptors such as post-synaptic AMPA receptors and pre-synaptic amyloid precursor protein. In addition, the variation in domain composition of these protein tools provides a means to dissect the Olfm1 regions important for receptor binding.http://link.springer.com/article/10.1186/s12860-019-0232-1Nervous systemSignallingSynapseProtein purificationX-ray crystallographySmall-angle X-ray scattering
collection DOAJ
language English
format Article
sources DOAJ
author Matti F. Pronker
Hugo van den Hoek
Bert J. C. Janssen
spellingShingle Matti F. Pronker
Hugo van den Hoek
Bert J. C. Janssen
Design and structural characterisation of olfactomedin-1 variants as tools for functional studies
BMC Molecular and Cell Biology
Nervous system
Signalling
Synapse
Protein purification
X-ray crystallography
Small-angle X-ray scattering
author_facet Matti F. Pronker
Hugo van den Hoek
Bert J. C. Janssen
author_sort Matti F. Pronker
title Design and structural characterisation of olfactomedin-1 variants as tools for functional studies
title_short Design and structural characterisation of olfactomedin-1 variants as tools for functional studies
title_full Design and structural characterisation of olfactomedin-1 variants as tools for functional studies
title_fullStr Design and structural characterisation of olfactomedin-1 variants as tools for functional studies
title_full_unstemmed Design and structural characterisation of olfactomedin-1 variants as tools for functional studies
title_sort design and structural characterisation of olfactomedin-1 variants as tools for functional studies
publisher BMC
series BMC Molecular and Cell Biology
issn 2661-8850
publishDate 2019-11-01
description Abstract Background Olfactomedin-1 (Olfm1; also known as Noelin or Pancortin) is a highly-expressed secreted brain and retina protein and its four isoforms have different roles in nervous system development and function. Structural studies showed that the long Olfm1 isoform BMZ forms a disulfide-linked tetramer with a V-shaped architecture. The tips of the Olfm1 “V” each consist of two C-terminal β-propeller domains that enclose a calcium binding site. Functional characterisation of Olfm1 may be aided by new biochemical tools derived from these core structural elements. Results Here we present the production, purification and structural analysis of three novel monomeric, dimeric and tetrameric forms of mammalian Olfm1 for functional studies. We characterise these constructs structurally by high-resolution X-ray crystallography and small-angle X-ray scattering. The crystal structure of the Olfm1 β-propeller domain (to 1.25 Å) represents the highest-resolution structure of an olfactomedin family member to date, revealing features such as a hydrophilic tunnel containing water molecules running into the core of the domain where the calcium binding site resides. The shorter Olfactomedin-1 isoform BMY is a disulfide-linked tetramer with a shape similar to the corresponding region in the longer BMZ isoform. Conclusions These recombinantly-expressed protein tools should assist future studies, for example of biophysical, electrophysiological or morphological nature, to help elucidate the functions of Olfm1 in the mature mammalian brain. The control over the oligomeric state of Olfm1 provides a firm basis to better understand the role of Olfm1 in the (trans-synaptic) tethering or avidity-mediated clustering of synaptic receptors such as post-synaptic AMPA receptors and pre-synaptic amyloid precursor protein. In addition, the variation in domain composition of these protein tools provides a means to dissect the Olfm1 regions important for receptor binding.
topic Nervous system
Signalling
Synapse
Protein purification
X-ray crystallography
Small-angle X-ray scattering
url http://link.springer.com/article/10.1186/s12860-019-0232-1
work_keys_str_mv AT mattifpronker designandstructuralcharacterisationofolfactomedin1variantsastoolsforfunctionalstudies
AT hugovandenhoek designandstructuralcharacterisationofolfactomedin1variantsastoolsforfunctionalstudies
AT bertjcjanssen designandstructuralcharacterisationofolfactomedin1variantsastoolsforfunctionalstudies
_version_ 1724443472407035904