# Accumulation of storage proteins in plant seeds is mediated by amyloid formation

**Authors:** Kirill S. Antonets, Mikhail V. Belousov, Anna I. Sulatskaya, Maria E. Belousova, Anastasiia O. Kosolapova, Maksim I. Sulatsky, Elena A. Andreeva, Pavel A. Zykin, Yury V. Malovichko, Oksana Y. Shtark, Anna N. Lykholay, Kirill V. Volkov, Irina M. Kuznetsova, Konstantin K. Turoverov, Elena Y. Kochetkova, Alexander G. Bobylev, Konstantin S. Usachev, Oleg. N. Demidov, Igor A. Tikhonovich, Anton A. Nizhnikov, Lauren A Richardson, Lauren A Richardson, Ines Alvarez-Garcia, Ines Alvarez-Garcia

PMC · DOI: 10.1371/journal.pbio.3000564 · PLoS Biology · 2020-07-23

## TL;DR

This study shows that storage proteins in garden pea seeds form stable amyloid structures, which resist digestion and canning.

## Contribution

The first demonstration of functional amyloid formation in plant seeds under native conditions.

## Key findings

- Vicilin, a storage protein in peas, forms amyloid fibrils in vivo and in vitro.
- Vicilin amyloids resist digestion and canning, and are toxic to yeast and mammalian cells.
- Amyloid formation of Vicilin increases during seed maturation and decreases during germination.

## Abstract

Amyloids are protein aggregates with a highly ordered spatial structure giving them unique physicochemical properties. Different amyloids not only participate in the development of numerous incurable diseases but control vital functions in archaea, bacteria and eukarya. Plants are a poorly studied systematic group in the field of amyloid biology. Amyloid properties have not yet been demonstrated for plant proteins under native conditions in vivo. Here we show that seeds of garden pea Pisum sativum L. contain amyloid-like aggregates of storage proteins, the most abundant one, 7S globulin Vicilin, forms bona fide amyloids in vivo and in vitro. Full-length Vicilin contains 2 evolutionary conserved β-barrel domains, Cupin-1.1 and Cupin-1.2, that self-assemble in vitro into amyloid fibrils with similar physicochemical properties. However, Cupin-1.2 fibrils unlike Cupin-1.1 can seed Vicilin fibrillation. In vivo, Vicilin forms amyloids in the cotyledon cells that bind amyloid-specific dyes and possess resistance to detergents and proteases. The Vicilin amyloid accumulation increases during seed maturation and wanes at germination. Amyloids of Vicilin resist digestion by gastrointestinal enzymes, persist in canned peas, and exhibit toxicity for yeast and mammalian cells. Our finding for the first time reveals involvement of amyloid formation in the accumulation of storage proteins in plant seeds.

Amyloids are protein fibrils that may be pathological or functional. This study of garden peas shows that several storage proteins accumulate in plant seeds as highly stable amyloids that resist the canning process and digestion by gastrointestinal enzymes.

## Linked entities

- **Proteins:** vicilin (vicilin), cupin1.2 (cupin superfamily member 1 gene 2)

## Full-text entities

- **Genes:** LDHC (lactate dehydrogenase C) [NCBI Gene 3948] {aka CT32, LDH3, LDHX}, SNCA (synuclein alpha) [NCBI Gene 6622] {aka NACP, PARK1, PARK4, PD1}, HTT (huntingtin) [NCBI Gene 3064] {aka HD, IT15, LOMARS}, SUP35 (translation termination factor GTPase eRF3) [NCBI Gene 851752] {aka GST1, PNM2, SAL3, SUF12, SUP2, SUP36}, APP (amyloid beta precursor protein) [NCBI Gene 351] {aka AAA, ABETA, ABPP, AD1, APPI, CTFgamma}, INS (insulin) [NCBI Gene 3630] {aka IDDM, IDDM1, IDDM2, ILPR, IRDN, MODY10}, Vicilin [NCBI Gene 103453212], CUP1-1 (metallothionein CUP1-1) [NCBI Gene 856450] {aka CUP1}, SAL1 (Ca(2+)-binding ATP:ADP antiporter SAL1) [NCBI Gene 855641], CDC39 (CCR4-NOT core subunit CDC39) [NCBI Gene 850455] {aka NOT1, ROS1, SMD6}, LYZ (lysozyme) [NCBI Gene 4069] {aka AMYLD5, LYZF1, LZM}, PGA5 (pepsinogen A5) [NCBI Gene 5222] {aka Pg5}
- **Diseases:** dehydration (MESH:D003681), necrosis (MESH:D009336), amyloid formation (MESH:D058426), amyloid fibrils (MESH:D014693), colorectal adenocarcinoma (MESH:D003110), amyloidoses (MESH:D000686), Amyloid (MESH:C000718787), food allergy (MESH:D005512), cytotoxic (MESH:D064420), Alzheimer disease (MESH:D000544), Amyloid deposition (MESH:D058225), neurodegenerative disorders (MESH:D019636)
- **Species:** Saccharomyces cerevisiae (baker's yeast, species) [taxon 4932], Homo sapiens (human, species) [taxon 9606], Powellomyces sp. EA (species) [taxon 252690], Rhizobium leguminosarum (species) [taxon 384], Austrofundulus limnaeus (species) [taxon 52670], Zea mays (maize, species) [taxon 4577], Escherichia coli (E. coli, species) [taxon 562], Escherichia coli BL21 (strain) [taxon 511693], Lathyrus oleraceus (garden pea, species) [taxon 3888], Mus musculus (house mouse, species) [taxon 10090], Oryctolagus cuniculus (domestic rabbit, species) [taxon 9986]
- **Cell lines:** DH5alpha — Drosophila hydei (Fruit fly), Spontaneously immortalized cell line (CVCL_Z531), S2 — Drosophila melanogaster (Fruit fly), Spontaneously immortalized cell line (CVCL_Z232), S2E — Mus musculus (Mouse), Hybridoma (CVCL_C5DX), PB — Homo sapiens (Human), Human papillomavirus-related endocervical adenocarcinoma, Cancer cell line (CVCL_JA22), DLD1 — Homo sapiens (Human), Colon adenocarcinoma, Cancer cell line (CVCL_0248)

## Full text

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## Figures

6 figures with captions in the complete paper: https://tomesphere.com/paper/PMC7377382/full.md

## References

104 references — full list in the complete paper: https://tomesphere.com/paper/PMC7377382/full.md

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Source: https://tomesphere.com/paper/PMC7377382