Large shifts in pKa values of lysine residues buried inside a protein

Daniel Isom, Carlos A. Castañed, Brian R. Cannon, Bertrand E. García-Moreno

Research output: Contribution to journalArticle

208 Citations (Scopus)

Abstract

Internal ionizable groups in proteins are relatively rare but they are essential for catalysis and energy transduction. To examine molecular determinants of their unusual and functionally important properties, we engineered 25 variants of staphylococcal nuclease with lysine residues at internal positions. Nineteen of the Lys residues have depressed pKa values, some as low as 5.3, and 20 titrate without triggering any detectable conformational reorganization. Apparently, simply by being buried in the protein interior, these Lys residues acquired pKa values comparable to those of naturally occurring internal ionizable groups involved in catalysis and biological H+ transport. The pKa values of some of the internal Lys residues were affected by interactions with surface carboxylic groups. The apparent polarizability reported by the pKa values varied significantly from location to location inside the protein. These data will enable an unprecedented examination of the positional dependence of the dielectric response of a protein. This study also shows that the ability of proteins to withstand the presence of charges in their hydrophobic interior is a fundamental property inherent to all stable proteins, not a specialized adaptation unique to proteins that evolved to depend on internal charges for function.

Original languageEnglish (US)
Pages (from-to)5260-5265
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume108
Issue number13
DOIs
StatePublished - Mar 29 2011
Externally publishedYes

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Lysine
Proteins
Catalysis
Biological Transport
Micrococcal Nuclease

ASJC Scopus subject areas

  • General

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Large shifts in pKa values of lysine residues buried inside a protein. / Isom, Daniel; Castañed, Carlos A.; Cannon, Brian R.; García-Moreno, Bertrand E.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 108, No. 13, 29.03.2011, p. 5260-5265.

Research output: Contribution to journalArticle

Isom, Daniel ; Castañed, Carlos A. ; Cannon, Brian R. ; García-Moreno, Bertrand E. / Large shifts in pKa values of lysine residues buried inside a protein. In: Proceedings of the National Academy of Sciences of the United States of America. 2011 ; Vol. 108, No. 13. pp. 5260-5265.
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