Investigation of dynamic solution interactions between NET-1 and UNC-5B by multi-wavelength analytical ultracentrifugation

IF 2.2 4区 生物学 Q3 BIOPHYSICS European Biophysics Journal Pub Date : 2023-03-20 DOI:10.1007/s00249-023-01644-1
Haben Gabir, Monika Gupta, Markus Meier, Fabian Heide, Manuel Koch, Joerg Stetefeld, Borries Demeler
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引用次数: 2

Abstract

NET-1 is a key chemotropic ligand that signals commissural axon migration and change in direction. NET-1 and its receptor UNC-5B switch axon growth cones from attraction to repulsion. The biophysical properties of the NET-1 + UNC-5B complex have been poorly characterized. Using multi-wavelength-AUC by adding a fluorophore to UNC-5B, we were able to separate the UNC-5B sedimentation from NET-1. Using both multi-wavelength- and single-wavelength AUC, we investigated NET-1 and UNC-5B hydrodynamic parameters and complex formation. The sedimentation velocity experiments show that NET-1 exists in a monomer–dimer equilibrium. A close study of the association shows that NET-1 forms a pH-sensitive dimer that interacts in an anti-parallel orientation. UNC-5B can form equimolar NET-1 + UNC-5B heterocomplexes with both monomeric and dimeric NET-1.

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NET-1与UNC-5B动态溶液相互作用的多波长超离心分析研究
NET-1是一种关键的趋化配体,它指示交联轴突的迁移和方向改变。NET-1及其受体UNC-5B将轴突生长锥从吸引转向排斥。NET-1 + UNC-5B络合物的生物物理性质尚不清楚。通过在UNC-5B上添加荧光团,使用多波长auc,我们能够从NET-1中分离UNC-5B沉淀物。使用多波长和单波长AUC,我们研究了NET-1和UNC-5B的水动力参数和复杂地层。沉降速度实验表明,NET-1存在于单体-二聚体平衡态。对这种关联的密切研究表明,NET-1形成了一种ph敏感二聚体,以反平行方向相互作用。UNC-5B可以与单体和二聚体NET-1形成等摩尔的NET-1 + UNC-5B异质配合物。
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来源期刊
European Biophysics Journal
European Biophysics Journal 生物-生物物理
CiteScore
4.30
自引率
0.00%
发文量
43
审稿时长
6-12 weeks
期刊介绍: The journal publishes papers in the field of biophysics, which is defined as the study of biological phenomena by using physical methods and concepts. Original papers, reviews and Biophysics letters are published. The primary goal of this journal is to advance the understanding of biological structure and function by application of the principles of physical science, and by presenting the work in a biophysical context. Papers employing a distinctively biophysical approach at all levels of biological organisation will be considered, as will both experimental and theoretical studies. The criteria for acceptance are scientific content, originality and relevance to biological systems of current interest and importance. Principal areas of interest include: - Structure and dynamics of biological macromolecules - Membrane biophysics and ion channels - Cell biophysics and organisation - Macromolecular assemblies - Biophysical methods and instrumentation - Advanced microscopics - System dynamics.
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