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Molecular crowding impacts the structure of apolipoprotein A-I with potential implications on in vivo metabolism and function

Petrlova, Jitka LU ; Hilt, Silvia; Budamagunta, Madhu; Domingo-Espín, Joan LU ; Voss, John C. LU and Lagerstedt, Jens O. LU (2016) In Biopolymers 105(10). p.683-692
Abstract

The effect molecular crowding, defined as the volume exclusion exerted by one soluble inert molecule upon another soluble molecule, has on the structure and self-interaction of lipid-free apoA-I were explored. The influence of molecular crowding on lipid-free apoA-I oligomerization and internal dynamics has been analyzed using electron paramagnetic resonance (EPR) spectroscopy measurements of nitroxide spin label at selected positions throughout the protein sequence and at varying concentrations of the crowding agent Ficoll-70. The targeted positions include sites previously shown to be sensitive for detecting intermolecular interaction via spin–spin coupling. Circular dichroism was used to study secondary structural changes in... (More)

The effect molecular crowding, defined as the volume exclusion exerted by one soluble inert molecule upon another soluble molecule, has on the structure and self-interaction of lipid-free apoA-I were explored. The influence of molecular crowding on lipid-free apoA-I oligomerization and internal dynamics has been analyzed using electron paramagnetic resonance (EPR) spectroscopy measurements of nitroxide spin label at selected positions throughout the protein sequence and at varying concentrations of the crowding agent Ficoll-70. The targeted positions include sites previously shown to be sensitive for detecting intermolecular interaction via spin–spin coupling. Circular dichroism was used to study secondary structural changes in lipid-free apoA-I imposed by increasing concentrations of the crowding agent. Crosslinking and SDS-PAGE gel analysis was employed to further characterize the role molecular crowding plays in inducing apoA-I oligomerization. It was concluded that the dynamic apoA-I structure and oligomeric state was altered in the presence of the crowding agent. It was also found that the C-terminal was slightly more sensitive to molecular crowding. Finally, the data described the region around residue 217 in the C-terminal domain of apoA-I as the most sensitive reporter of the crowding-induced self-association of apoA-I. The implications of this behavior to in vivo functionality are discussed. © 2016 Wiley Periodicals, Inc. Biopolymers 105: 683–692, 2016.

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
apoA-I, apolipoprotein A-I, CD, EPR, Ficoll-70, HDL, high-density lipoprotein, molecular crowding
in
Biopolymers
volume
105
issue
10
pages
10 pages
publisher
John Wiley & Sons
external identifiers
  • Scopus:84978971672
  • WOS:000383369400003
ISSN
0006-3525
DOI
10.1002/bip.22865
language
English
LU publication?
yes
id
1d5b329f-05c2-43d7-bf6e-dfb7135fb69e
date added to LUP
2016-10-17 10:26:21
date last changed
2017-01-01 08:37:00
@article{1d5b329f-05c2-43d7-bf6e-dfb7135fb69e,
  abstract     = {<p>The effect molecular crowding, defined as the volume exclusion exerted by one soluble inert molecule upon another soluble molecule, has on the structure and self-interaction of lipid-free apoA-I were explored. The influence of molecular crowding on lipid-free apoA-I oligomerization and internal dynamics has been analyzed using electron paramagnetic resonance (EPR) spectroscopy measurements of nitroxide spin label at selected positions throughout the protein sequence and at varying concentrations of the crowding agent Ficoll-70. The targeted positions include sites previously shown to be sensitive for detecting intermolecular interaction via spin–spin coupling. Circular dichroism was used to study secondary structural changes in lipid-free apoA-I imposed by increasing concentrations of the crowding agent. Crosslinking and SDS-PAGE gel analysis was employed to further characterize the role molecular crowding plays in inducing apoA-I oligomerization. It was concluded that the dynamic apoA-I structure and oligomeric state was altered in the presence of the crowding agent. It was also found that the C-terminal was slightly more sensitive to molecular crowding. Finally, the data described the region around residue 217 in the C-terminal domain of apoA-I as the most sensitive reporter of the crowding-induced self-association of apoA-I. The implications of this behavior to in vivo functionality are discussed. © 2016 Wiley Periodicals, Inc. Biopolymers 105: 683–692, 2016.</p>},
  author       = {Petrlova, Jitka and Hilt, Silvia and Budamagunta, Madhu and Domingo-Espín, Joan and Voss, John C. and Lagerstedt, Jens O.},
  issn         = {0006-3525},
  keyword      = {apoA-I,apolipoprotein A-I,CD,EPR,Ficoll-70,HDL,high-density lipoprotein,molecular crowding},
  language     = {eng},
  month        = {10},
  number       = {10},
  pages        = {683--692},
  publisher    = {John Wiley & Sons},
  series       = {Biopolymers},
  title        = {Molecular crowding impacts the structure of apolipoprotein A-I with potential implications on in vivo metabolism and function},
  url          = {http://dx.doi.org/10.1002/bip.22865},
  volume       = {105},
  year         = {2016},
}