NMR Determination of Protein pK(a) Values in the Solid State

Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801.
Journal of Physical Chemistry Letters (Impact Factor: 7.46). 05/2010; 1(10):1623-1628. DOI: 10.1021/jz1004413
Source: PubMed

ABSTRACT Charged residues play an important role in defining key mechanistic features in many biomolecules. Determining the pK(a) values of large, membrane or fibrillar proteins can be challenging with traditional methods. In this study we show how solid-state NMR is used to monitor chemical shift changes during a pH titration for the small soluble β1 immunoglobulin binding domain of protein G. The chemical shifts of all the amino acids with charged side-chains throughout the uniformly-(13)C,(15)N-labeled protein were monitored over several samples varying in pH; pK(a) values were determined from these shifts for E27, D36, and E42, and the bounds for the pK(a) of other acidic side-chain resonances were determined. Additionally, this study shows how the calculated pK(a) values give insights into the crystal packing of the protein.

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