Acid-induced denaturation of myoglobin studied by time-resolved electrospray ionization mass spectrometry

Biochemistry
L KonermannD J Douglas

Abstract

The acid-induced denaturation of holo-myoglobin (hMb) following a pH-jump from 6.5 to 3.2 has been studied by electrospray ionization (ESI) mass spectrometry in combination with a continuous flow mixing technique (time-resolved ESI MS). Different protein conformations are detected by the different charge state distributions that they generate during ESI. The changes in intensity of the peaks in the mass spectrum as a function of time can be described by two exponential lifetimes of 0.38 +/- 0.06 s and 6.1 +/- 0.5 s, respectively. The acid-induced denaturation of hMb was also studied in stopped-flow experiments by monitoring changes in the Soret absorption. The lifetimes measured by this method are in good agreement with those obtained by time-resolved ESI MS. The shorter lifetime is associated with the formation of a transient intermediate which shows the mass of the intact heme-protein complex but leads to the formation of much higher charge states during ESI than native hMb at pH 6.5. This form of hMb has an absorption spectrum similar to that of the native protein, indicating a relatively unperturbed chromophore environment inside the heme binding pocket. The intermediate can thus be characterized as an unfolded form of hMb ...Continue Reading

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Citations

May 29, 2000·Rapid Communications in Mass Spectrometry : RCM·B M KolakowskiL Konermann
May 28, 2003·Mass Spectrometry Reviews·Lars Konermann, Douglas A Simmons
Feb 27, 2001·Rapid Communications in Mass Spectrometry : RCM·B B SchneiderD D Chen
Dec 18, 2001·Rapid Communications in Mass Spectrometry : RCM·P A ChrismanS A McLuckey
Dec 18, 2001·Rapid Communications in Mass Spectrometry : RCM·H Lin, C Dass
Dec 10, 2003·Analytical and Bioanalytical Chemistry·Maria SamalikovaRita Grandori
Sep 29, 2011·Journal of the American Society for Mass Spectrometry·Rui WangRenato Zenobi
Sep 29, 2011·Journal of the American Society for Mass Spectrometry·David J ClarkePat R R Langridge-Smith
Sep 29, 2011·Journal of the American Society for Mass Spectrometry·Atim A EnyenihiRoman A Zubarev
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May 24, 2005·Journal of the American Society for Mass Spectrometry·Atsushi TakamizawaKenzo Hiraoka
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