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Pregled bibliografske jedinice broj: 1229180

Heterologous expression and purification of SH2 domain-containing protein 3, protein with a predicted high content of intrinsically disordered regions


Matovina, Mihaela; Tomašić Paić, Ana; Matić, Sara; Barbarić, Lea; Crnolatac, Ivo; Berger, Tamara; Miočić Stošić, Fran
Heterologous expression and purification of SH2 domain-containing protein 3, protein with a predicted high content of intrinsically disordered regions // FEBS Open Bio Volume 12 Supplement 1 : The Biochemistry Global Summit, 25th IUBMB Congress, 46th FEBS Congress, 15th PABMB Congres
Lisabon, Portugal, 2022. str. 258-258 (poster, podatak o recenziji nije dostupan, sažetak, znanstveni)


CROSBI ID: 1229180 Za ispravke kontaktirajte CROSBI podršku putem web obrasca

Naslov
Heterologous expression and purification of SH2 domain-containing protein 3, protein with a predicted high content of intrinsically disordered regions

Autori
Matovina, Mihaela ; Tomašić Paić, Ana ; Matić, Sara ; Barbarić, Lea ; Crnolatac, Ivo ; Berger, Tamara ; Miočić Stošić, Fran

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

Izvornik
FEBS Open Bio Volume 12 Supplement 1 : The Biochemistry Global Summit, 25th IUBMB Congress, 46th FEBS Congress, 15th PABMB Congres / - , 2022, 258-258

Skup
The Biochemistry Global Summit

Mjesto i datum
Lisabon, Portugal, 06.07.2022. - 14.07.2022

Vrsta sudjelovanja
Poster

Vrsta recenzije
Podatak o recenziji nije dostupan

Ključne riječi
SH2D3C ; protein expression ; internally disordered regions (IDRs)

Sažetak
SH2 domain-containing protein 3C (SH2D3C) is one of only three members of the family of proteins that contain both SH2 domain and a domain similar to guanine nucleotide exchange factor domains for Ras family GTPases (Ras GEF- like domain). It acts as an adapter protein and has proposed roles in cell migration and adhesion, tissue organization, and the regulation of the immune response. The data about the structure of the SH2D3C protein are scarce. Only one structure can be found in the Protein Data Bank, and it is the structure of the C-terminal, Ras GEF-like domain in the complex with the Breast cancer anti-estrogen resistance protein 1 (BCAR1). SH2D3C protein was identified as a prey in the SILAC-MS analysis of the dipeptidyl peptidase 3 (DPP3) interactome. To prepare the purified protein for the biochemical and biophysical analysis of SH2D3C and its interaction with DPP3, we have expressed isoforms 2 and 3 of SH2D3C and the C-terminal domain in E. coli with HIS-, GST- and MBP-tag. We were able to purify only the C-terminal domain with GST- and MBP-tag, but the proteolytic removal of the tag rendered the domain unstable, however, isoform 3 was successfully expressed and purified by baculovirus-mediated protein expression in the insect cells. Isoform 3 lacks the N-terminal SH2 domain and has only the Ras GEF-like domain. According to the AlphaFold prediction of the structure of the longest, isoform 1 which contains both SH2 and Ras GEF-like domains, a large part of isoform 3 should be disordered. This was confirmed by CD measurements of the purified isoform 3 which showed that around 43 % of the protein is disordered. SEC analysis of isoform 3 indicated that it forms dimers, however, SEC-MALS showed that it is a monomer, which also indicates that the protein is probably largely disordered. Despite having a high proportion of internally disordered regions, SH2D3C-isoform 3 is stable and will be used for further biochemical, biophysical and structural analysis.

Izvorni jezik
Engleski

Znanstvena područja
Kemija, Biologija, Interdisciplinarne prirodne znanosti



POVEZANOST RADA


Projekti:
--IP-2020-02-6743 - Interakcija dipeptidil peptidaze III s proteinom SH2 domain-containing protein 3C – moguća veza između odgovora na oksidativni stres i stanične migracije (OxMiLink) (Matovina, Mihaela) ( CroRIS)

Ustanove:
Institut "Ruđer Bošković", Zagreb


Citiraj ovu publikaciju:

Matovina, Mihaela; Tomašić Paić, Ana; Matić, Sara; Barbarić, Lea; Crnolatac, Ivo; Berger, Tamara; Miočić Stošić, Fran
Heterologous expression and purification of SH2 domain-containing protein 3, protein with a predicted high content of intrinsically disordered regions // FEBS Open Bio Volume 12 Supplement 1 : The Biochemistry Global Summit, 25th IUBMB Congress, 46th FEBS Congress, 15th PABMB Congres
Lisabon, Portugal, 2022. str. 258-258 (poster, podatak o recenziji nije dostupan, sažetak, znanstveni)
Matovina, M., Tomašić Paić, A., Matić, S., Barbarić, L., Crnolatac, I., Berger, T. & Miočić Stošić, F. (2022) Heterologous expression and purification of SH2 domain-containing protein 3, protein with a predicted high content of intrinsically disordered regions. U: FEBS Open Bio Volume 12 Supplement 1 : The Biochemistry Global Summit, 25th IUBMB Congress, 46th FEBS Congress, 15th PABMB Congres.
@article{article, author = {Matovina, Mihaela and Toma\v{s}i\'{c} Pai\'{c}, Ana and Mati\'{c}, Sara and Barbari\'{c}, Lea and Crnolatac, Ivo and Berger, Tamara and Mio\v{c}i\'{c} Sto\v{s}i\'{c}, Fran}, year = {2022}, pages = {258-258}, keywords = {SH2D3C, protein expression, internally disordered regions (IDRs)}, title = {Heterologous expression and purification of SH2 domain-containing protein 3, protein with a predicted high content of intrinsically disordered regions}, keyword = {SH2D3C, protein expression, internally disordered regions (IDRs)}, publisherplace = {Lisabon, Portugal} }
@article{article, author = {Matovina, Mihaela and Toma\v{s}i\'{c} Pai\'{c}, Ana and Mati\'{c}, Sara and Barbari\'{c}, Lea and Crnolatac, Ivo and Berger, Tamara and Mio\v{c}i\'{c} Sto\v{s}i\'{c}, Fran}, year = {2022}, pages = {258-258}, keywords = {SH2D3C, protein expression, internally disordered regions (IDRs)}, title = {Heterologous expression and purification of SH2 domain-containing protein 3, protein with a predicted high content of intrinsically disordered regions}, keyword = {SH2D3C, protein expression, internally disordered regions (IDRs)}, publisherplace = {Lisabon, Portugal} }

Časopis indeksira:


  • Web of Science Core Collection (WoSCC)
    • Science Citation Index Expanded (SCI-EXP)
    • SCI-EXP, SSCI i/ili A&HCI
  • Scopus
  • MEDLINE





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