Journal article
Cytochrome c unfolding on an anionic surface
Journal of Chromatography A, Vol.863(2), pp.137-146
1999
Handle:
https://hdl.handle.net/2376/116978
PMID: 10593494
Abstract
It is now well accepted that the adsorption of proteins to solid supports sometimes involves surface-mediated unfolding. A detailed understanding of the adsorption and surface-mediated unfolding process is lacking. We selected a well studied protein, horse heart cytochrome
c, and a weakly ionic support to examine some of the characteristics of protein adsorption under near-physiological conditions. We used high-performance liquid chromatography (HPLC) to investigate the effect of temperature on surface-mediated unfolding. Samples of cytochrome
c were introduced to an anionic support, and a NaCl gradient was used to desorb the protein at different times and temperatures. The profiles and retention times were monitored to examine the adhesive properties of cytochrome
c to the anionic support. We found that protein retention increased with time at temperatures as low as 0°C, and a significant loss of cytochrome
c occurred between 55°C and 70°C. The loss of recovery of cytochrome
c indicates irreversible surface-mediated unfolding. The changes in retention time may indicate more subtle transitions, including reversible surface-mediated unfolding of cytochrome
c . These results suggest that perturbations in the structure as well as unfolding of cytochrome
c can be detected at a lower temperature on an anionic surface than in solution thereby acting like a catalyst for protein unfolding.
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Details
- Title
- Cytochrome c unfolding on an anionic surface
- Creators
- Craig W HerboldJohn H MillerSteven C Goheen
- Publication Details
- Journal of Chromatography A, Vol.863(2), pp.137-146
- Academic Unit
- Engineering and Applied Sciences (TRIC), School of
- Publisher
- Elsevier B.V
- Identifiers
- 99900547526901842
- Language
- English
- Resource Type
- Journal article