skip to main content

DOE PAGESDOE PAGES

Title: One Crystal, Two Temperatures: Cryocooling Penalties Alter Ligand Binding to Transient Protein Sites

Interrogating fragment libraries by X-ray crystallography is a powerful strategy for discovering allosteric ligands for protein targets. Cryocooling of crystals should theoretically increase the fraction of occupied binding sites and decrease radiation damage. However, it might also perturb protein conformations that can be accessed at room temperature. Using data from crystals measured consecutively at room temperature and at cryogenic temperature, we found that transient binding sites could be abolished at the cryogenic temperatures employed by standard approaches. Finally, changing the temperature at which the crystallographic data was collected could provide a deliberate perturbation to the equilibrium of protein conformations and help to visualize hidden sites with great potential to allosterically modulate protein function.
Authors:
 [1] ;  [1] ;  [2]
  1. Univ. of California, San Francisco, CA (United States). Dept. of Pharmaceutical Chemistry
  2. Univ. of California, San Francisco, CA (United States). Dept. of Bioengineering and Therapeutic Sciences
Publication Date:
Grant/Contract Number:
AC02-05CH11231; GM59957; OD009180; GM110580; STC-1231306
Type:
Accepted Manuscript
Journal Name:
ChemBioChem: a European journal of chemical biology
Additional Journal Information:
Journal Volume: 16; Journal Issue: 11; Journal ID: ISSN 1439-4227
Publisher:
ChemPubSoc Europe
Research Org:
Univ. of California, San Francisco, CA (United States)
Sponsoring Org:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); National Institutes of Health (NIH)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 36 MATERIALS SCIENCE; allosterism; biophysics; ligand discovery; structural biology; thermodynamics; X-ray diffraction
OSTI Identifier:
1343096