We studied the effects of humic substances (HS) extracted from soil on the identification of the recombinant ovine prion protein (RecPrP) by denaturing (sodium dodecyl sulfate polyacrylamide gel electrophoresis [SDS-PAGE]) and native PAGE (N-PAGE), and mass spectrometry (MS), at various HS to RecPrP contact ratios. The results showed that the contact with HS did not alter RecPrP electrophoretic mobility but affected protein identification by MS. Contact between RecPrP and HS resulted in a lower coverage percentage of specific RecPrP domains that led to a prion misidentification, more evident after N-PAGE than SDS-PAGE. The analysis of the nonidentified protein domains suggests that lower quality of RecPrP identification could be due to hydrophobic interactions between the prion protein and HS, but the mechanism by which HS hamper the correct identification of RecPrP remains to be established. Our results may have implications in the prion environmental risk assessment.

Contact with soil-borne humic substances interfere with the prion identification by mass spectrometry

2014

Abstract

We studied the effects of humic substances (HS) extracted from soil on the identification of the recombinant ovine prion protein (RecPrP) by denaturing (sodium dodecyl sulfate polyacrylamide gel electrophoresis [SDS-PAGE]) and native PAGE (N-PAGE), and mass spectrometry (MS), at various HS to RecPrP contact ratios. The results showed that the contact with HS did not alter RecPrP electrophoretic mobility but affected protein identification by MS. Contact between RecPrP and HS resulted in a lower coverage percentage of specific RecPrP domains that led to a prion misidentification, more evident after N-PAGE than SDS-PAGE. The analysis of the nonidentified protein domains suggests that lower quality of RecPrP identification could be due to hydrophobic interactions between the prion protein and HS, but the mechanism by which HS hamper the correct identification of RecPrP remains to be established. Our results may have implications in the prion environmental risk assessment.
2014
Istituto di Ricerca sugli Ecosistemi Terrestri - IRET
RecPrP
Prion proteins
Humic substances
Prion interactions
Prion identification
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/20.500.14243/295233
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