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

Title: Engineered stabilization and structural analysis of the autoinhibited conformation of PDE4

Abstract

Phosphodiesterase 4 (PDE4) is an essential contributor to intracellular signaling and an important drug target. The four members of this enzyme family (PDE4A to -D) are functional dimers in which each subunit contains two upstream conserved regions (UCR), UCR1 and -2, which precede the C-terminal catalytic domain. Alternative promoters, transcriptional start sites, and mRNA splicing lead to the existence of over 25 variants of PDE4, broadly classified as long, short, and supershort forms. We report the X-ray crystal structure of long form PDE4B containing UCR1, UCR2, and the catalytic domain, crystallized as a dimer in which a disulfide bond cross-links cysteines engineered into UCR2 and the catalytic domain. Biochemical and mass spectrometric analyses showed that the UCR2-catalytic domain interaction occurs in trans, and established that this interaction regulates the catalytic activity of PDE4. By elucidating the key structural determinants of dimerization, we show that only long forms of PDE4 can be regulated by this mechanism. The results also provide a structural basis for the long-standing observation of high- and low-affinity binding sites for the prototypic inhibitor rolipram.

Authors:
 [1];  [1];  [1];  [1]; ORCiD logo [1]
  1. Pfizer, Inc., Groton, CT (United States). Structural Biology and Biophysics Group
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
INDUSTRY
OSTI Identifier:
1182326
Grant/Contract Number:  
AC02-06CH11357
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 112; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences, Washington, DC (United States)
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Cedervall, Peder, Aulabaugh, Ann, Geoghegan, Kieran F., McLellan, Thomas J., and Pandit, Jayvardhan. Engineered stabilization and structural analysis of the autoinhibited conformation of PDE4. United States: N. p., 2015. Web. doi:10.1073/pnas.1419906112.
Cedervall, Peder, Aulabaugh, Ann, Geoghegan, Kieran F., McLellan, Thomas J., & Pandit, Jayvardhan. Engineered stabilization and structural analysis of the autoinhibited conformation of PDE4. United States. doi:10.1073/pnas.1419906112.
Cedervall, Peder, Aulabaugh, Ann, Geoghegan, Kieran F., McLellan, Thomas J., and Pandit, Jayvardhan. Mon . "Engineered stabilization and structural analysis of the autoinhibited conformation of PDE4". United States. doi:10.1073/pnas.1419906112. https://www.osti.gov/servlets/purl/1182326.
@article{osti_1182326,
title = {Engineered stabilization and structural analysis of the autoinhibited conformation of PDE4},
author = {Cedervall, Peder and Aulabaugh, Ann and Geoghegan, Kieran F. and McLellan, Thomas J. and Pandit, Jayvardhan},
abstractNote = {Phosphodiesterase 4 (PDE4) is an essential contributor to intracellular signaling and an important drug target. The four members of this enzyme family (PDE4A to -D) are functional dimers in which each subunit contains two upstream conserved regions (UCR), UCR1 and -2, which precede the C-terminal catalytic domain. Alternative promoters, transcriptional start sites, and mRNA splicing lead to the existence of over 25 variants of PDE4, broadly classified as long, short, and supershort forms. We report the X-ray crystal structure of long form PDE4B containing UCR1, UCR2, and the catalytic domain, crystallized as a dimer in which a disulfide bond cross-links cysteines engineered into UCR2 and the catalytic domain. Biochemical and mass spectrometric analyses showed that the UCR2-catalytic domain interaction occurs in trans, and established that this interaction regulates the catalytic activity of PDE4. By elucidating the key structural determinants of dimerization, we show that only long forms of PDE4 can be regulated by this mechanism. The results also provide a structural basis for the long-standing observation of high- and low-affinity binding sites for the prototypic inhibitor rolipram.},
doi = {10.1073/pnas.1419906112},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = ,
volume = 112,
place = {United States},
year = {Mon Mar 09 00:00:00 EDT 2015},
month = {Mon Mar 09 00:00:00 EDT 2015}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Citation Metrics:
Cited by: 18 works
Citation information provided by
Web of Science

Save / Share: