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Native state dynamics affects the folding transition of porcine pancreatic phospholipase A2.

Kölbel, Knut; Weininger, Ulrich LU ; Ihling, Christian; Mrestani-Klaus, Carmen and Ulbrich-Hofmann, Renate (2015) In Biophysical Chemistry 206. p.12-21
Abstract
Porcine pancreatic phospholipase A2, a small and disulfide rich protein, is extremely resistant against chemically or thermally induced unfolding. Despite this marked resistance, the protein displays broad unfolding transitions resulting in comparatively low apparent thermodynamic stability. Broad unfolding transitions may result from undetected folding intermediates, residual structures in the unfolded state or an inhomogeneity of the native state. Using circular dichroism, fluorescence, and NMR spectroscopy, we ruled out the existence of stably populated folding intermediates, whereas UV absorbance measurements hinted at stable residual structures in the unfolded state. These residual structures proved, however, to have no impact on the... (More)
Porcine pancreatic phospholipase A2, a small and disulfide rich protein, is extremely resistant against chemically or thermally induced unfolding. Despite this marked resistance, the protein displays broad unfolding transitions resulting in comparatively low apparent thermodynamic stability. Broad unfolding transitions may result from undetected folding intermediates, residual structures in the unfolded state or an inhomogeneity of the native state. Using circular dichroism, fluorescence, and NMR spectroscopy, we ruled out the existence of stably populated folding intermediates, whereas UV absorbance measurements hinted at stable residual structures in the unfolded state. These residual structures proved, however, to have no impact on the folding parameters. Studies by limited proteolysis, CD, and NMR spectroscopy under non-denaturing conditions suggested pronounced dynamics of the protein in the native state, which as long as unrestrained by acidic pH or bound Ca(2+) ions exert considerable influence on the unfolding transition. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Biophysical Chemistry
volume
206
pages
12 - 21
publisher
Elsevier
external identifiers
  • pmid:26117657
  • wos:000361259000002
  • scopus:84934922295
ISSN
1873-4200
DOI
10.1016/j.bpc.2015.06.007
language
English
LU publication?
yes
id
d1bf2220-1df8-4698-b96c-90c26b880481 (old id 7477634)
date added to LUP
2015-07-17 09:35:43
date last changed
2017-01-01 04:05:40
@article{d1bf2220-1df8-4698-b96c-90c26b880481,
  abstract     = {Porcine pancreatic phospholipase A2, a small and disulfide rich protein, is extremely resistant against chemically or thermally induced unfolding. Despite this marked resistance, the protein displays broad unfolding transitions resulting in comparatively low apparent thermodynamic stability. Broad unfolding transitions may result from undetected folding intermediates, residual structures in the unfolded state or an inhomogeneity of the native state. Using circular dichroism, fluorescence, and NMR spectroscopy, we ruled out the existence of stably populated folding intermediates, whereas UV absorbance measurements hinted at stable residual structures in the unfolded state. These residual structures proved, however, to have no impact on the folding parameters. Studies by limited proteolysis, CD, and NMR spectroscopy under non-denaturing conditions suggested pronounced dynamics of the protein in the native state, which as long as unrestrained by acidic pH or bound Ca(2+) ions exert considerable influence on the unfolding transition.},
  author       = {Kölbel, Knut and Weininger, Ulrich and Ihling, Christian and Mrestani-Klaus, Carmen and Ulbrich-Hofmann, Renate},
  issn         = {1873-4200},
  language     = {eng},
  pages        = {12--21},
  publisher    = {Elsevier},
  series       = {Biophysical Chemistry},
  title        = {Native state dynamics affects the folding transition of porcine pancreatic phospholipase A2.},
  url          = {http://dx.doi.org/10.1016/j.bpc.2015.06.007},
  volume       = {206},
  year         = {2015},
}