Naslov
Aggregation of the protein TRIOBP-1, involved in schizophrenia, depends on two of its structural regions

Autori
Hart, Anja ; Odorčić, Maja ; Zaharija, Beti ; Bradshaw, Nicholas J.

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Book of Abstracts, 29th International Student Congress of (bio)Medical Sciences / - , 2022, 369-369

Skup
29th International Student Congress of (bio)Medical Sciences

Mjesto i datum
Groningen, Nizozemska, 08.06.2022. - 10.06.2022

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
Schizophrenia ; Protein aggregation ; TRIOBP-1

Sažetak
Introduction Schizophrenia is a severe mental illness, characterized by life-long cognitive and behavioural changes. Its development is caused by genetic and environmental factors whose complexity has made it difficult to invent new treatment methods. It has recently been suggested that aberrant proteostasis may also impact the progression of schizophrenia. If certain misfolded proteins, caused by the disruption in proteostasis, are not degraded by the cell, aggregation occurs and the protein forms large insoluble structures known as aggregates. TRIO and F-actin binding protein (TRIOBP-1) is one such protein which aggregates in a subgroup of patients. It has previously been shown that aggregation of TRIOBP-1 may depend on a region in the centre of the protein, but this has only been shown using plasmids encoding parts of the protein. Materials & Methods PCR was used to clone different plasmid constructs, encoding full length or partial TRIOBP-1, fused with FLAG-tag, which were then propagated in NEB5α bacteria. Purified plasmids were transfected in HEK239 human kidney cells and expression confirmed by Western blotting. Constructs were then overexpressed in SH-SY5Y neuroblastoma cells to visualise aggregation using immunofluorescent microscopy. All experiments and results were repeated 3 times and verified using negative controls. Results Constructs encoding TRIOBP-1 lacking different versions of the aggregation-critical region still aggregated when overexpressed in SH-SY5Y, suggesting that another structural part is also involved in TRIOBP-1’s aggregation propensity. Based on previous results and research, we hypothesised that the optionally translated N- terminus of TRIOBP-1 might also influence aggregation. We confirmed this by truncating the full-length construct further, to exclude the N- terminal unstructured region. All these results lead to generating a non-aggregating TRIOBP-1 mutant (60-652, Δ333-340). Conclusion We have shown two regions to be responsible for the aggregation propensity of TRIOBP-1: the optionally translated N-terminus and amino acids 333-340 in the central section of the protein. By generating mutant TRIOBP-1 with the minimal number of mutations required to prevent aggregation we will be able to generate model systems for studying TRIOBP-1 aggregation, and allow us to better understand its role in the progression of schizophrenia.

Izvorni jezik
Engleski

Znanstvena područja
Biotehnologija u biomedicini (prirodno područje, biomedicina i zdravstvo, biotehničko područje)

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