The influence of glycation on a high pressure denaturation of ubiquitin.
The combination of deuterium-hydrogen exchange (DHX) and mass spectrometry (MS) can be used for studying a high pressure denaturation (HPD) of proteins. Herein we present the results of investigations of the influence of glycation on the HPD of ubiquitin. Application of various values of pressure causes different degrees of protein unfolding, resulting in molecules with a different number of protons available for exchange with deuterons. The dependence of this number on pressure gives information on the denaturation state of a protein. On the basis of the obtained results we can conclude that increasing number of fructosamine moieties in ubiquitin decreases the pressure required for its denaturation. It suggests that glycation moderately decreases the protein stability. The present study is the first example of application of hydrogen-deuterium exchange as a method of investigating the influence of posttranslational modification of protein on the HPD.
Faculty of Chemistry, University of Wrocław, Wrocław 50-383, Poland monika.kijewska@chem.uni.wroc.pl.
Monika
M
Kijewska
Faculty of Chemistry, University of Wrocław, Wrocław 50-383, Poland.
Karolina
K
Radziszewska
Faculty of Chemistry, University of Wrocław, Wrocław 50-383, Poland.
Marta
M
Cal
Faculty of Chemistry, University of Wrocław, Wrocław 50-383, Poland.
Mateusz
M
Waliczek
Faculty of Chemistry, University of Wrocław, Wrocław 50-383, Poland.
Piotr
P
Stefanowicz
Faculty of Chemistry, University of Wrocław, Wrocław 50-383, Poland.
Zbigniew
Z
Szewczuk
D003903:Deuterium / Q000737:chemistry; D041961:Deuterium Exchange Measurement / Q000379:methods; D006031:Glycosylation; D006859:Hydrogen / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods; D011312:Pressure; D011489:Protein Denaturation; D017510:Protein Folding; D055550:Protein Stability; D025801:Ubiquitin / Q000737:chemistry*
D025801:Ubiquitin; D006859:Hydrogen; D003903:Deuterium
high-pressure denaturation; hydrogen–deuterium exchange; mass spectrometry; non-enzymatic glycation