التفاصيل البيبلوغرافية
| العنوان: |
Biochemical and Biophysical Characterization of the Caveolin-2 Interaction with Membranes and Analysis of the Protein Structural Alteration by the Presence of Cholesterol |
| المؤلفون: |
Gorospe, Berta, Moura, José J. G., Gutierrez-Merino, Carlos, Samhan-Arias, Alejandro K. |
| المساهمون: |
DQ - Departamento de Química, LAQV@REQUIMTE, RUN |
| سنة النشر: |
2022 |
| وصف مادي: |
18 |
| مصطلحات موضوعية: |
caveolin-1, caveolin-2, cholesterol, membrane interaction, secondary structure prediction, Catalysis, Molecular Biology, Spectroscopy, Computer Science Applications, Physical and Theoretical Chemistry, Organic Chemistry, Inorganic Chemistry |
| الوصف: |
This work was supported by the Associate Laboratory for Green Chemistry—LAQV which is financed by national funds from FCT/MCTES (UID/QUI/50006/2020) and co-financed by the ERDF under the PT2020 Partnership Agreement (POCI-01-0145-FEDER—007265). Experimental work was also supported by funding from Ayuda a Grupos de la Junta de Extremadura (Group BBB008-GR21051), co-financed by the European Funds for Structural Development (FEDER). The BioLab is co-financed by the ERDF under the PT2020 Partnership Agreement (POCI-01-0145-FEDER-007728 and POCI-01-0145-FEDER—007265, respectively). Publisher Copyright: © 2022 by the authors. |
| الوصف (مترجم): |
Caveolin-2 is a protein suitable for the study of interactions of caveolins with other proteins and lipids present in caveolar lipid rafts. Caveolin-2 has a lower tendency to associate with high molecular weight oligomers than caveolin-1, facilitating the study of its structural modulation upon association with other proteins or lipids. In this paper, we have successfully expressed and purified recombinant human caveolin-2 using E. coli. The structural changes of caveolin-2 upon interaction with a lipid bilayer of liposomes were characterized using bioinformatic prediction models, circular dichroism, differential scanning calorimetry, and fluorescence techniques. Our data support that caveolin-2 binds and alters cholesterol-rich domains in the membranes through a CARC domain, a type of cholesterol-interacting domain in its sequence. The far UV-CD spectra support that the purified protein keeps its folding properties but undergoes a change in its secondary structure in the presence of lipids that correlates with the acquisition of a more stable conformation, as shown by differential scanning calorimetry experiments. Fluorescence experiments using egg yolk lecithin large unilamellar vesicles loaded with 1,6-diphenylhexatriene confirmed that caveolin-2 adsorbs to the membrane but only penetrates the core of the phospholipid bilayer if vesicles are supplemented with 30% of cholesterol. Our study sheds light on the caveolin-2 interaction with lipids. In addition, we propose that purified recombinant caveolin-2 can provide a new tool to study protein–lipid interactions within caveolae. |
| نوع الوثيقة: |
journal article |
| وصف الملف: |
application/pdf |
| اللغة: |
English |
| العلاقة: |
Gorospe, B., Moura, J. J. G., Gutierrez-Merino, C., & Samhan-Arias, A. K. (2022). Biochemical and Biophysical Characterization of the Caveolin-2 Interaction with Membranes and Analysis of the Protein Structural Alteration by the Presence of Cholesterol. International Journal of Molecular Sciences, 23(23), [15203]. https://doi.org/10.3390/ijms232315203Test; 1661-6596; PURE: 55548660 |
| DOI: |
10.3390/ijms232315203 |
| الإتاحة: |
http://hdl.handle.net/10362/150485Test |
| حقوق: |
open access |
| رقم الانضمام: |
rcaap.com.unl.run.unl.pt.10362.150485 |
| قاعدة البيانات: |
RCAAP |
