Journal article
Hydrogen Bonding Markedly Reduces the p K of Buried Carboxyl Groups in Proteins
Journal of molecular biology, Vol.362(3), pp.594-604
2006
DOI: 10.1016/j.jmb.2006.07.056
Abstract
The ionizable groups in proteins with the lowest p
Ks are the carboxyl groups of aspartic acid side-chains. One of the lowest, p
K
=
0.6, is observed for Asp76 in ribonuclease T1. This low p
K appeared to result from hydrogen bonds to a water molecule and to the side-chains of Asn9, Tyr11, and Thr91. The results here confirm this by showing that the p
K of Asp76 increases to 1.7 in N9A, to 4.0 in Y11F, to 4.2 in T91V, to 4.4 in N9A
+
Y11F, to 4.9 in N9A
+
T91V, to 5.9 in Y11F
+
T91V, and to 6.4 in the triple mutant: N9A
+
Y11F
+
T91V. In ribonuclease Sa, the lowest p
K
=
2.4 for Asp33. This p
K increases to 3.9 in T56A, which removes the hydrogen bond to Asp33, and to 4.4 in T56V, which removes the hydrogen bond and replaces the –OH group with a –CH
3 group. It is clear that hydrogen bonds are able to markedly lower the p
K values of carboxyl groups in proteins. These same hydrogen bonds make large contributions to the conformational stability of the proteins. At pH 7, the stability of D76A ribonuclease T1 is 3.8 kcal mol
−1 less than wild-type, and the stability of D33A ribonuclease Sa is 4.1 kcal mol
−1 less than wild-type. There is a good correlation between the changes in the p
K values and the changes in stability. The results suggest that the p
K values for these buried carboxyl groups would be greater than 8 in the absence of hydrogen bonds, and that the hydrogen bonds and other interactions of the carboxyl groups contribute over 8 kcal mol
−1 to the stability.
Details
- Title: Subtitle
- Hydrogen Bonding Markedly Reduces the p K of Buried Carboxyl Groups in Proteins
- Creators
- Richard L. Thurlkill - Texas A&M University SystemGerald R. Grimsley - Texas A&M University SystemJ. Martin Scholtz - Texas A&M UniversityC. Nick Pace - Texas A&M University
- Resource Type
- Journal article
- Publication Details
- Journal of molecular biology, Vol.362(3), pp.594-604
- Publisher
- Elsevier Ltd
- DOI
- 10.1016/j.jmb.2006.07.056
- ISSN
- 0022-2836
- eISSN
- 1089-8638
- Language
- English
- Date published
- 2006
- Academic Unit
- Research Administration; Pharmaceutical Sciences and Experimental Therapeutics; Biochemistry and Molecular Biology; Chemistry
- Record Identifier
- 9984288715802771
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