Thermal stability analyses of human PERIOD-2 C-terminal domain using dynamic light scattering and circular dichroism

Autor: Noah J. Ruelas, Karla Moriel, Paulina Rios, Neha Chauhan, Seham Z. Azzam, Yuejiao Xian, Jennie Choi, Ruben Cano Rodriguez, Brenda Moreno, Victoria Miranda, Bianca N. Amezaga, Luis C. Nunez, Adan E. Castaneda, Christian S. Quinones, Chuan Xiao, Neha Vijay
Jazyk: angličtina
Rok vydání: 2020
Předmět:
Models
Molecular
Circular dichroism
Protein Folding
Circadian clock
Thermal Stability
Biochemistry
Protein Structure
Secondary
Database and Informatics Methods
0302 clinical medicine
Protein structure
Post-Translational Modification
Materials
0303 health sciences
Multidisciplinary
Crystallography
Chemistry
Protein Stability
Physics
Circular Dichroism
Temperature
Classical Mechanics
Period Circadian Proteins
Condensed Matter Physics
PER2
Circadian Rhythms
Physical Sciences
Crystal Structure
Thermodynamics
Disulfide Bonds
Medicine
Protein folding
Sequence Analysis
Research Article
Bioinformatics
Period (gene)
Science
Protein domain
Materials Science
Fluid Mechanics
Research and Analysis Methods
Continuum Mechanics
03 medical and health sciences
Protein Domains
Solid State Physics
Humans
Amino Acid Sequence
Dimers
030304 developmental biology
C-terminus
Biology and Life Sciences
Proteins
Fluid Dynamics
Polymer Chemistry
Dynamic Light Scattering
Oligomers
Biophysics
Hydrodynamics
Sequence Alignment
Chronobiology
030217 neurology & neurosurgery
Zdroj: PLoS ONE, Vol 15, Iss 4, p e0221180 (2020)
PLoS ONE
ISSN: 1932-6203
Popis: At the molecular level, the circadian clock is regulated by a time delayed transcriptional-translational feedback loop in which the core proteins interact with each other rhythmically to drive daily biological rhythms. The C-terminal domain of a key clock protein PER2 (PER2c) plays a critically important role in the loop, not only for its interaction with the binding partner CRY proteins but also for the CRY/PER complex's translocation from the cytosol to the nucleus. Previous circular dichroism (CD) spectroscopic studies have shown that mouse PER2c (mPER2c) is less structured in solution by itself but folded into stable secondary structures upon interaction with mouse CRYs. To understand the stability and folding of human PER2c (hPER2c), we expressed and purified hPER2c. Three oligomerization forms of recombinant hPER2c were identified and thoroughly characterized through a combination of biochemical and biophysical techniques. Different to mPER2c, both thermal unfolding DLS and CD analyses suggested that all forms of hPER2c have very stable secondary structures in solution by themselves with melting temperatures higher than the physiological body temperature, indicating that hPER2c does not require CRY to fold. Furthermore, we examined the effects of EDTA, salt concentration, and a reducing agent on hPER2c folding and oligomerization. The ability of hPER2c forming oligomers reflects the potential role of hPER2c in the assembly of circadian rhythm core protein complexes.
Databáze: OpenAIRE
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