Selective isolation of hydrophobin SC3 by solid-phase extraction with polytetrafluoroethylene microparticles and subsequent mass spectrometric analysis
Language English Country Germany Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
- Keywords
- Hydrophobin SC3, Isolation, Mass spectrometry, Polytetrafluoroethylene microparticles, Protein analysis,
- MeSH
- Albumins chemistry MeSH
- Ananas chemistry MeSH
- Bromelains chemistry MeSH
- Canavalia chemistry MeSH
- Chymotrypsin chemistry MeSH
- Cytochromes c chemistry MeSH
- Disulfides chemistry MeSH
- Erythrocytes enzymology MeSH
- Solid Phase Extraction methods MeSH
- Mass Spectrometry methods MeSH
- Carbonic Anhydrases chemistry MeSH
- Caseins chemistry MeSH
- Horses MeSH
- Concanavalin A chemistry MeSH
- Chickens MeSH
- Humans MeSH
- Microspheres * MeSH
- Milk enzymology MeSH
- Myocardium metabolism MeSH
- Polytetrafluoroethylene chemistry MeSH
- Proteomics methods MeSH
- Schizophyllum chemistry MeSH
- Cattle MeSH
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization MeSH
- Tandem Mass Spectrometry MeSH
- Thermolysin chemistry MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Cattle MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Albumins MeSH
- Bromelains MeSH
- Chymotrypsin MeSH
- Cytochromes c MeSH
- Disulfides MeSH
- Carbonic Anhydrases MeSH
- Caseins MeSH
- Concanavalin A MeSH
- Polytetrafluoroethylene MeSH
- Thermolysin MeSH
Hydrophobins are small proteins that play a role in a number of processes during the filamentous fungi growth and development. These proteins are characterized by the self-assembly of their molecules into an amphipathic membrane at hydrophilic-hydrophobic interfaces. Isolation and purification of hydrophobins generally present a challenge in their analysis. Hydrophobin SC3 from Schizophyllum commune was selected as a representative of class I hydrophobins in this work. A novel procedure for selective and effective isolation of hydrophobin SC3 based on solid-phase extraction with polytetrafluoroethylene microparticles loaded in a small self-made microcolumn is reported. The tailored binding of hydrophobins to polytetrafluoroethylene followed by harsh elution conditions resulted in a highly specific isolation of hydrophobin SC3 from the model mixture of ten proteins. The presented isolation protocol can have a positive impact on the analysis and utilization of these proteins including all class I hydrophobins. Hydrophobin SC3 was further subjected to reduction of its highly stable disulfide bonds and to chymotryptic digestion followed by mass spectrometric analysis. The isolation and digestion protocols presented in this work make the analysis of these highly hydrophobic and compact proteins possible.
References provided by Crossref.org
HU protein is involved in intracellular growth and full virulence of Francisella tularensis
