skip to main content
DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Extensive site-directed mutagenesis reveals interconnected functional units in the alkaline phosphatase active site

Abstract

Enzymes enable life by accelerating reaction rates to biological timescales. Conventional studies have focused on identifying the residues that have a direct involvement in an enzymatic reaction, but these so-called ‘catalytic residues’ are embedded in extensive interaction networks. Although fundamental to our understanding of enzyme function, evolution, and engineering, the properties of these networks have yet to be quantitatively and systematically explored. We dissected an interaction network of five residues in the active site of Escherichia coli alkaline phosphatase. Analysis of the complex catalytic interdependence of specific residues identified three energetically independent but structurally interconnected functional units with distinct modes of cooperativity. From an evolutionary perspective, this network is orders of magnitude more probable to arise than a fully cooperative network. From a functional perspective, new catalytic insights emerge. Further, such comprehensive energetic characterization will be necessary to benchmark the algorithms required to rationally engineer highly efficient enzymes.

Authors:
 [1];  [1];  [1];  [2];  [1]
  1. Department of Biochemistry, Beckman Center, Stanford University, Stanford, United States
  2. Molecular and Cellular Biochemistry Department, Indiana University Bloomington, Bloomington, United States
Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
1182427
Alternate Identifier(s):
OSTI ID: 1198402
Grant/Contract Number:  
Office of Biological and Environmental Research
Resource Type:
Published Article
Journal Name:
eLife
Additional Journal Information:
Journal Name: eLife Journal Volume: 4; Journal ID: ISSN 2050-084X
Publisher:
eLife Sciences Publications, Ltd.
Country of Publication:
United States
Language:
English

Citation Formats

Sunden, Fanny, Peck, Ariana, Salzman, Julia, Ressl, Susanne, and Herschlag, Daniel. Extensive site-directed mutagenesis reveals interconnected functional units in the alkaline phosphatase active site. United States: N. p., 2015. Web. doi:10.7554/eLife.06181.
Sunden, Fanny, Peck, Ariana, Salzman, Julia, Ressl, Susanne, & Herschlag, Daniel. Extensive site-directed mutagenesis reveals interconnected functional units in the alkaline phosphatase active site. United States. doi:10.7554/eLife.06181.
Sunden, Fanny, Peck, Ariana, Salzman, Julia, Ressl, Susanne, and Herschlag, Daniel. Wed . "Extensive site-directed mutagenesis reveals interconnected functional units in the alkaline phosphatase active site". United States. doi:10.7554/eLife.06181.
@article{osti_1182427,
title = {Extensive site-directed mutagenesis reveals interconnected functional units in the alkaline phosphatase active site},
author = {Sunden, Fanny and Peck, Ariana and Salzman, Julia and Ressl, Susanne and Herschlag, Daniel},
abstractNote = {Enzymes enable life by accelerating reaction rates to biological timescales. Conventional studies have focused on identifying the residues that have a direct involvement in an enzymatic reaction, but these so-called ‘catalytic residues’ are embedded in extensive interaction networks. Although fundamental to our understanding of enzyme function, evolution, and engineering, the properties of these networks have yet to be quantitatively and systematically explored. We dissected an interaction network of five residues in the active site of Escherichia coli alkaline phosphatase. Analysis of the complex catalytic interdependence of specific residues identified three energetically independent but structurally interconnected functional units with distinct modes of cooperativity. From an evolutionary perspective, this network is orders of magnitude more probable to arise than a fully cooperative network. From a functional perspective, new catalytic insights emerge. Further, such comprehensive energetic characterization will be necessary to benchmark the algorithms required to rationally engineer highly efficient enzymes.},
doi = {10.7554/eLife.06181},
journal = {eLife},
number = ,
volume = 4,
place = {United States},
year = {2015},
month = {4}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
DOI: 10.7554/eLife.06181

Citation Metrics:
Cited by: 16 works
Citation information provided by
Web of Science

Save / Share:

Works referenced in this record:

Kemp elimination catalysts by computational enzyme design
journal, March 2008

  • Röthlisberger, Daniela; Khersonsky, Olga; Wollacott, Andrew M.
  • Nature, Vol. 453, Issue 7192, p. 190-195
  • DOI: 10.1038/nature06879

The role of induced fit and conformational changes of enzymes in specificity and catalysis
journal, March 1988