Projekteja vuodessa
Abstrakti
There is an increasing understanding that condensation is a crucial intermediate step in the assembly of biological materials and for a multitude of cellular processes. To apply and to understand these mechanisms, in vitro biophysical characterisation techniques are central. The formation and biophysical properties of protein condensates depend on a multitude of factors, such as protein concentration, pH, temperature, salt concentration, and presence of other biomolecules as well as protein purification and storage conditions. Here we show how critical the procedures for preparing protein samples for in vitro studies are. We compare two purification methods of the recombinant spider silk protein CBM-AQ12-CBM and study the effect of background molecules, such as DNA, on the formation and properties of the condensates. We characterize the condensates using aggregation induced emitters (AIEs), coalescence studies, and micropipette aspiration. The condensated sample containing background molecules exhibit a lower threshold concentration for condensate formation accompanied by a lower surface tension and longer coalescence time when compared to the pure protein condensates. Furthermore, the partitioning of small AIEs is enhanced in the presence of background molecules. Our results highlight that the purification method and remaining background molecules strongly affect the biophysical properties of spider silk condensates. Using the acquired knowledge about spider silk protein purification we derive guidelines for reproducible condensate formation that will foster the use of spider silk proteins as adhesives or carriers for biomedical applications.
Alkuperäiskieli | Englanti |
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Sivut | 11953-11967 |
Sivumäärä | 15 |
Julkaisu | Journal of Materials Chemistry B |
Vuosikerta | 12 |
Numero | 46 |
Varhainen verkossa julkaisun päivämäärä | 16 lokak. 2024 |
DOI - pysyväislinkit | |
Tila | Julkaistu - 14 jouluk. 2024 |
OKM-julkaisutyyppi | A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä |
Sormenjälki
Sukella tutkimusaiheisiin 'Recombinant silk protein condensates show widely different properties depending on the sample background'. Ne muodostavat yhdessä ainutlaatuisen sormenjäljen.Tietoaineistot
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Data for the publication: Recombinant silk protein condensates show widely different properties depending on the sample background
Tersteegen, J. (Creator), Tunn, I. (Creator), Sand, M. (Creator), Välisalmi, T. (Creator), Malkamäki, M. (Creator), Gandier, J.-A. (Creator), Beaune, G. (Creator), Sanz Velasco, A. (Creator), Anaya-Plaza, E. (Creator) & Linder, M. B. (Creator), Zenodo, 16 lokak. 2024
DOI - pysyväislinkki: 10.5281/zenodo.12529261, https://zenodo.org/records/12529262
Tietoaineisto: Dataset
Projektit
- 5 Aktiivinen
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-: LIBER Linder II
Linder, M. (Vastuullinen tutkija), Yin, Y. (Projektin jäsen), Malkamäki, M. (Projektin jäsen) & Fedorov, D. (Projektin jäsen)
01/01/2024 → 31/12/2026
Projekti: Academy of Finland: Other research funding
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LIBER Linder: Life-like hybrid materials
Linder, M. (Vastuullinen tutkija), Elfving, K. (Projektin jäsen), Lemetti, L. (Projektin jäsen), Tunn, I. (Projektin jäsen), Aspelin, H. (Projektin jäsen), Roas Escalona, N. (Projektin jäsen), Aranko, S. (Projektin jäsen), Malkamäki, M. (Projektin jäsen), Osmekhina, E. (Projektin jäsen) & Fedorov, D. (Projektin jäsen)
01/01/2022 → 31/12/2026
Projekti: Academy of Finland: Other research funding
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PhotoCage: Photoactive protein cage biohybrids for light-induced applications
Anaya, E. (Vastuullinen tutkija)
01/09/2021 → 31/08/2026
Projekti: Academy of Finland: Other research funding
Laitteet
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Biotalousinfrastruktuuri
Seppälä, J. (Manager)
Kemian tekniikan korkeakouluLaitteistot/tilat: Facility
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OtaNano Nanomikroskopiakeskus
Seitsonen, J. (Manager) & Rissanen, A. (Other)
OtaNanoLaitteistot/tilat: Facility