Abstract
The light chain of the major histocompatibility complex class 1 (MHC-1), the protein β 2-microglobulin (β 2m), has amyloidogenic properties that arise only upon its dissociation from the MHC-1. Here hydrogen/deuterium exchange electrospray ionization mass spectrometry (HDX-ESI-MS) has been used to compare the solution dynamics of β 2m in its MHC-1 bound state compared with those of β 2m as a free monomer. The capability of tandem mass spectrometry to dissociate the MHC-1 into its individual constituents in the gas phase following deuterium incorporation in solution has permitted the direct observation of the exchange properties of MHC-1 bound β 2m for the first time. The HDX-ESI-MS data show clearly that the H→D exchange of MHC-1 bound β 2m follows EX2 kinetics and that about 20 protons remain protected from exchange after 17 days. Free from the MHC-1, monomeric β 2m exhibits significantly different HDX behavior, which encompasses both EX1 and EX2 kinetics. The EX2 kinetics indicate a tenfold increase in the rate of exchange compared with MHC-1 bound β 2m, with just 10 protons remaining protected from EX2 exchange and therefore exchanging only via the EX1 mechanism. The EX1 kinetics observed for unbound β 2m are consistent with unfolding of its exchange-protected core with a t1/2 of 68 min (pH 7, 37° C). Thus, upon dissociation from the stabilizing influence of the MHC-1, free β 2m becomes highly dynamic and undergoes unfolding transitions that result in an aggregation-competent protein.
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Published online October 17, 2008
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Hodkinson, J.P., Jahn, T.R., Radford, S.E. et al. HDX-ESI-MS reveals enhanced conformational dynamics of the amyloidogenic protein β 2-microglobulin upon release from the MHC-1. J Am Soc Mass Spectrom 20, 278–286 (2009). https://doi.org/10.1016/j.jasms.2008.10.005
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DOI: https://doi.org/10.1016/j.jasms.2008.10.005