Fast Mapping of Global Protein Folding States by Multivariate NMR: A GPS for Proteins

Malmendal, Anders; Underhaug, Jarl; Otzen, Daniel E.; Nielsen, Niels Chr.

doi:10.1371/journal.pone.0010262

Cited by 17 publications

(21 citation statements)

References 34 publications

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“…Although it has been reported that low concentrations of SDS can induce cooperative conformational transitions, 7,28 this data shows that the effect of dodine is fundamentally different from the effect of its two constituent parts at the same concentration.…”

Section: Resultscontrasting

confidence: 57%

Dodine as a Protein Denaturant: The Best of Two Worlds?

2013

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Traditional denaturants such as urea and guanidinium ion unfold proteins in a cooperative “all-or-none” fashion. However, their high working concentration in combination with their strong absorption in the far ultraviolet region make it impossible to measure high quality circular dichroism or infrared spectra, which are commonly used to detect changes in protein secondary structure. On the other hand, detergents such as dodecyl sulfate destabilize native protein conformation at low millimolar concentrations and are UV transparent, but they do denature proteins more gradually than guanidinium or urea. In this work we studied the denaturation properties of the fungicide dodecylguanidinium acetate (dodine), which combines both denaturants into one. We show that dodine unfolds some small proteins at millimolar concentrations, facilitates temperature denaturation, and is transparent enough at its working concentration, unlike guanidinium, to measure full range circular dichroism spectra. Our results also suggest that dodine allows fine-tuning of the protein’s unfolded state, unlike traditional “all-or-none” denaturants.

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Section: Resultscontrasting

confidence: 57%

Dodine as a Protein Denaturant: The Best of Two Worlds?

2013

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“…A weakness with the chromatographic approach is that the method does not allow us to control the protein-lipid ratio and, indeed, it often leads to complexes with highly variable stoichiometries [7,[30][31][32]37]. Initial efforts to produce HAMLET complexes by other approaches such as simple mixing under different solvent conditions were not successful [2,33,38].…”

Section: Preparation Of Cytotoxic A-la:oa Complexes By Different Apprmentioning

confidence: 99%

Protein–fatty acid complexes: biochemistry, biophysics and function

2013

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Thirteen years ago, α‐lactalbumin (α‐LA) was first reported to form a complex with oleic acid (OA). This complex, called HAMLET (human α‐lactalbumin made lethal to tumour cells), was found to be cytotoxic to cancer cells. In HAMLET, α‐LA assumes a partially unfolded conformation and can bind OA in various stoichiometries. Subsequently, different groups have been able to prepare HAMLET‐like cytotoxic complexes in different ways, which all involve the destabilization of α‐LA, and a number of different proteins have been able to form similar complexes. This suggests that the ability to form stable complexes with lipids may be a generic feature of the polypeptide chain, although the precise structural and functional details may vary from protein to protein. We review the biophysical and biochemical properties of this class of complexes, focusing on different methods of preparation, complex structure and the role of the protein and the lipid within these complexes. The cellular effects of these complexes are multifaceted and depend on the cell types. There are strong indications that OA has an essential role, whereas the protein component, rather than having a toxic effect on its own, functions as a vehicle for transporting the toxic OA to the cells and keeping the OA in solution. Fatty acids alone can affect numerous cellular signalling and metabolic pathways, in addition to playing important roles in immune responses and inflammatory processes. Further studies will aim to determine how the molecular properties of the different protein–lipid complexes correlate with their biological efficacy.

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“…S6). A small cooperative transition in the signal with C m = 0.08 m M is followed by a gradual loss of structure as a linear function of SDS concentration.…”

Section: Resultsmentioning

confidence: 97%

Dodine as a transparent protein denaturant for circular dichroism and infrared studies

et al. 2016

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The fungicide dodine combines the cooperative denaturation properties of guanidine with the mM denaturation activity of SDS. It was previously tested only on two small model proteins. Here we show that it can be used as a chemical denaturant for phosphoglycerate kinase (PGK), a much larger two-domain enzyme. In addition to its properties as a chemical denaturant, dodine facilitates thermal denaturation of PGK, and we show for the first time that it also facilitates pressure denaturation of a protein. Much higher quality circular dichroism and amide I' infrared spectra of PGK can be obtained in dodine than in guanidine, opening the possibility for use of dodine as a denaturant when UV or IR detection is desirable. One caution is that dodine denaturation, like other detergent-based denaturants, is less reversible than guanidine denaturation.

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Fast Mapping of Global Protein Folding States by Multivariate NMR: A GPS for Proteins

Cited by 17 publications

References 34 publications

Dodine as a Protein Denaturant: The Best of Two Worlds?

Dodine as a Protein Denaturant: The Best of Two Worlds?

Protein–fatty acid complexes: biochemistry, biophysics and function

Dodine as a transparent protein denaturant for circular dichroism and infrared studies

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