Pregled bibliografske jedinice broj: 1228358
Mapping the Domain Structure and Aggregation Propensity of Proteins Using a Gateway Plasmid Vector System
Mapping the Domain Structure and Aggregation Propensity of Proteins Using a Gateway Plasmid Vector System // Protein aggregation: / Cieplak, Andrzej Stanisław (ur.).
New York (NY): Humana Press, 2023. str. 649-677 doi:10.1007/978-1-0716-2597-2_39
CROSBI ID: 1228358 Za ispravke kontaktirajte CROSBI podršku putem web obrasca
Naslov
Mapping the Domain Structure and Aggregation
Propensity of Proteins Using a Gateway Plasmid
Vector System
Autori
Zaharija, Beti ; Bradshaw, Nicholas J.
Vrsta, podvrsta i kategorija rada
Poglavlja u knjigama, znanstveni
Knjiga
Protein aggregation:
Urednik/ci
Cieplak, Andrzej Stanisław
Izdavač
Humana Press
Grad
New York (NY)
Godina
2023
Raspon stranica
649-677
ISBN
978-1-0716-2596-5
Ključne riječi
Gateway cloning ; Immunocytochemistry ; Immunofluorescent microscopy ; Protein aggregation ; Protein domains ; Protein folding ; Protein solubility ; Recombinant protein ; Size exclusion chromatography ; TRIOBP-1
Sažetak
Some proteins represent members of conserved families, meaning that their domain structure can be easily predicted by comparison to homologous proteins whose structures have been solved experimentally. Many other proteins, however, do not share significant detectable homology with other proteins, often as results of high amounts of coiled-coil structure and/or intrinsically unstructured regions. These proteins include many whose aggregation is linked to human disease. Here we present a refined and reliable workflow for identifying the domains of such proteins, through cloning of multiple alternative fragments, and testing whether they form soluble, folded structures when expressed as recombinant peptides in E. coli, through the use of size exclusion chromatography. By using Gateway recombination for cloning, these fragments can then be rapidly transferred to alternate vectors for testing in mammalian cells. We then specifically illustrate its use for proteins that form pathological aggregates in disease, mapping not just their basic domain structures but also the specific subdomains responsible for aggregation
Izvorni jezik
Engleski
Znanstvena područja
Biologija, Biotehnologija u biomedicini (prirodno područje, biomedicina i zdravstvo, biotehničko područje)
POVEZANOST RADA
Projekti:
--IP-2018-01-9424 - Istraživanje shizofrenije kroz ekspresiju netopivih proteina (CandidIskren) (Bradshaw, Nicholas James) ( CroRIS)
--DOK-2018-09-5395 - Istraživanje shizofrenije kroz ekspresiju netopivih proteina (CandidIskren) (Bradshaw, Nicholas James) ( CroRIS)
NadSve-Sveučilište u Rijeci-17.12.2.1.03 - Sumoilacija proteina uključenih u kronične mentalne bolesti (Bradshaw, Nicholas James, NadSve - Sveučilište u Rijeci - Inicijalne potpore mladim istraživačima 2017) ( CroRIS)
Ustanove:
Sveučilište u Rijeci - Odjel za biotehnologiju