Cysteine pKa depression by a protonated glutamic acid in human DJ-1

Anna C. Witt, Mahadevan Lakshminarasimhan, Benjamin C. Remington, Sahar Hasim, Edwin Pozharski, Mark A. Wilson

Research output: Contribution to journalArticle

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Abstract

Human DJ-1, a disease-associated protein that protects cells from oxidative stress, contains an oxidation-sensitive cysteine (C106) that is essential for its cytoprotective activity. The origin of C106 reactivity is obscure, due in part to the absence of an experimentally determined pKa value for this residue. We have used atomic-resolution X-ray crystallography and UV spectroscopy to show that C106 has a depressed pKa of 5.4 ± 0.1 and that the C106 thiolate accepts a hydrogen bond from a protonated glutamic acid side chain (E18). X-ray crystal structures and cysteine pK a analysis of several site-directed substitutions at residue 18 demonstrate that the protonated carboxylic acid side chain of E18 is required for the maximal stabilization of the C106 thiolate. A nearby arginine residue (R48) participates in a guanidinium stacking interaction with R28 from the other monomer in the DJ-1 dimer and elevates the pKa of C106 by binding an anion that electrostatically suppresses thiol ionization. Our results show that the ionizable residues (E18, R48, and R28) surrounding C106 affect its pK a in a way that is contrary to expectations based on the typical ionization behavior of glutamic acid and arginine. Lastly, a search of the Protein Data Bank (PDB) produces several candidate hydrogen-bonded aspartic/glutamic acid-cysteine interactions, which we propose are particularly common in the DJ-1 superfamily.

Original languageEnglish (US)
Pages (from-to)7430-7440
Number of pages11
JournalBiochemistry
Volume47
Issue number28
DOIs
StatePublished - Jul 15 2008

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Cysteine
Glutamic Acid
Ionization
Arginine
Hydrogen
Oxidative stress
X ray crystallography
X Ray Crystallography
Guanidine
Carboxylic Acids
Ultraviolet spectroscopy
Sulfhydryl Compounds
Dimers
Anions
Spectrum Analysis
Hydrogen bonds
Proteins
Oxidative Stress
Substitution reactions
Stabilization

ASJC Scopus subject areas

  • Biochemistry

Cite this

Witt, A. C., Lakshminarasimhan, M., Remington, B. C., Hasim, S., Pozharski, E., & Wilson, M. A. (2008). Cysteine pKa depression by a protonated glutamic acid in human DJ-1. Biochemistry, 47(28), 7430-7440. https://doi.org/10.1021/bi800282d

Cysteine pKa depression by a protonated glutamic acid in human DJ-1. / Witt, Anna C.; Lakshminarasimhan, Mahadevan; Remington, Benjamin C.; Hasim, Sahar; Pozharski, Edwin; Wilson, Mark A.

In: Biochemistry, Vol. 47, No. 28, 15.07.2008, p. 7430-7440.

Research output: Contribution to journalArticle

Witt, AC, Lakshminarasimhan, M, Remington, BC, Hasim, S, Pozharski, E & Wilson, MA 2008, 'Cysteine pKa depression by a protonated glutamic acid in human DJ-1', Biochemistry, vol. 47, no. 28, pp. 7430-7440. https://doi.org/10.1021/bi800282d
Witt AC, Lakshminarasimhan M, Remington BC, Hasim S, Pozharski E, Wilson MA. Cysteine pKa depression by a protonated glutamic acid in human DJ-1. Biochemistry. 2008 Jul 15;47(28):7430-7440. https://doi.org/10.1021/bi800282d
Witt, Anna C. ; Lakshminarasimhan, Mahadevan ; Remington, Benjamin C. ; Hasim, Sahar ; Pozharski, Edwin ; Wilson, Mark A. / Cysteine pKa depression by a protonated glutamic acid in human DJ-1. In: Biochemistry. 2008 ; Vol. 47, No. 28. pp. 7430-7440.
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