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Aggregation of alpha-synuclein protein in human blood plasma/serum

Aggregation of alpha-synuclein protein in human blood plasma/serum

BACHELOR project and MASTER project   (published Januari 2019)

Introduction

Parkinsons disease (PD), the second most frequently occurring neurodegenerative disease in the world, is mainly characterized by the loss of dopaminergic neurons and the presence of insoluble proteic complexes, so-called Lewy bodies (LB), in the brain [1]. LB are largely composed of fibrillar aggregates (amyloid) of the protein alpha-synuclein (aSyn), which suggests that aSyn plays an important role in the pathogenesis of PD. In addition, aSyn gene mutations and duplication/triplication are associated with hereditary forms of PD [2].

The aSyn protein is small (14 kDa), intrinsically disordered, and in vivo mainly expressed in dopaminergic neurons in the brain [3]. Several studies showed that aSyn is secreted by neuronal cells in the brain and ends up cerebrospinal fluid (CSF) and human blood plasma [5, 6]. In addition, a recent report demonstrated that aSyn can bi-directionally cross the blood brain barrier [7]. This implicates a potential risk to develop PD when people are in contact with aSyn species via their blood, which may happen through blood transfusion or accidentally in a laboratory setting. Potentially the foreign aSyn species may induce or seed (enhance) the aggregation of endogenous aSyn in the recipient brain. Therefore it is relevant to study the aSyn aggregation characteristics in body fluids, like human blood plasma/serum.

 Description of the project:

Testing the aggregation properties of aSyn in human blood serum.

Previously we have done some initial experiments to study the ability of aSyn to aggregate in serum. In these in vitro experiments we made use of recombinantly expressed and purified aSyn protein. We compared aSyn aggregation characteristics in buffer and in foetal calf serum.

The aim of this project is to investigate the aSyn aggregation properties in the more relevant human blood serum environment. By this way, we will see if the compounds present inside the human blood serum can modify (induction/inhibition) the aggregation characteristics of aSyn and so if aSyn can aggregate itself inside the human blood. To correctly investigate the effect of the blood human serum of the aSyn aggregation, the serum used will be initially purified of the native aSyn present.

 Techniques involved:

  • Affinity chromatography
  • In vitro aSyn aggregation reactions
  • Thioflavin T fluorescence measurements
  • Turbidity measurements
  •  Circular Dichroism
  • Atomic force microscopy

 

Contact:

Department: NanoBioPhysics (NBP) – UTwente - https://www.utwente.nl/en/tnw/nbp/

Daily supervisor: MSc. Jonathan Vaneyck (PhD student) - j.vaneyck@utwente.nl, Dr. G.M.J. Segers-Nolten (Researcher) - g.m.j.segers-nolten@utwente.nl

Project leader: Prof.Dr. M.M.A.E. Claessens (Chair Nanobiophysics) - m.m.a.e.claessens@utwente.nl