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Urea as a protein destabilizing agent in electrospray ionisation
Author(s) -
Donald Lynda J.,
Collado Vladimir M.,
Galka Jamie J.,
O'Neil Joe D.,
Duckworth Harry W.,
Loewen Peter C.,
Standing Kenneth G.
Publication year - 2009
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.3941
Subject(s) - chemistry , urea , electrospray ionization , non covalent interactions , mass spectrometry , electrospray , chromatography , crystallography , molecule , hydrogen bond , biochemistry , organic chemistry
Urea is well known as a denaturant of proteins, but there is also evidence that millimolar amounts of urea may in fact stabilize protein complexes. Advances in mass spectrometric analysis have given us the opportunity to test the effect of urea on several noncovalent complexes in buffered solutions. We expected to see lower charge states if folded proteins were more compact (and therefore more stable), and higher charge states if the proteins were denatured. We have found that mM urea interferes with some noncovalent interactions, and that the extent of interference depends on the specific protein complex. The difference seems to be related to the type of interactions, with weak ones, such as H‐bonds, more sensitive to urea. Examples show that a quick check with urea may give some insights into protein stability in the mass spectrometer. Copyright © 2009 John Wiley & Sons, Ltd.