Localized hydration in lyophilized myoglobin by hydrogen-deuterium exchange mass spectrometry. 1. Exchange mapping.

Molecular Pharmaceutics
Andreas M SophocleousE M Topp

Abstract

The local effects of hydration on myoglobin (Mb) in solid matrices containing mannitol or sucrose (1:1 w/w, protein:additive) were mapped using hydrogen-deuterium exchange with mass spectrometric analysis (HDX-MS) at 5 °C and compared to solution controls. Solid powders were exposed to D₂O(g) at controlled activity (a(w)) followed by reconstitution and analysis of the intact protein and peptides produced by pepsin digestion. HDX varied with matrix type, a(w), and position along the protein backbone. HDX was less in sucrose matrices than in mannitol matrices at all a(w) while the difference in solution was negligible. Differences in HDX in the two matrices were detectable despite similarities in their bulk water content. The extent of exchange in solids is proposed as a measure of the hydration of exchangeable amide groups, as well as protein conformation and dynamics; pepsin digestion allows these effects to be mapped with peptide-level resolution.

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Citations

Feb 12, 2014·Computers & Chemical Engineering·Brock C RoughtonKyle V Camarda
Feb 23, 2012·Molecular Pharmaceutics·Andreas M Sophocleous, Elizabeth M Topp
Jan 30, 2015·Journal of Pharmaceutical Sciences·Saradha Chandrasekhar, Elizabeth M Topp
Nov 15, 2014·Analytical Chemistry·Gregory F PirroneJohn R Engen
Apr 6, 2016·Journal of Pharmaceutical Sciences·Lavanya K IyerElizabeth M Topp
Oct 16, 2013·Molecular Pharmaceutics·Lavanya K IyerElizabeth M Topp
Dec 25, 2019·Pharmaceutical Research·Nathan E WilsonQi Tony Zhou
Jan 25, 2021·International Journal of Pharmaceutics·Rishabh TukraElizabeth M Topp
Jul 30, 2021·International Journal of Pharmaceutics·Tarun Tejasvi MutukuriQi Tony Zhou
Feb 6, 2019·Molecular Pharmaceutics·Yuan Chen, Elizabeth M Topp
Jun 28, 2019·Molecular Pharmaceutics·Rajashekar Kammari, Elizabeth M Topp
Aug 14, 2020·Molecular Pharmaceutics·Rajashekar Kammari, Elizabeth M Topp

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