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

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 ?1 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.

SUBMITTER: Schmidt HL

PROVIDER: S-EPMC2885713 | biostudies-literature | 2010 May

REPOSITORIES: biostudies-literature

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

Schmidt Heather L Frericks HL Shah Gautam J GJ Sperling Lindsay J LJ Rienstra Chad M CM

The journal of physical chemistry letters 20100501 10

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-labele ...[more]

PMID: 20563223

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