title: A linear relationship between crystal size and fragment binding time observed crystallographically: implications for fragment library screening using acoustic droplet ejection.
abstract: High throughput screening technologies such as acoustic droplet ejection (ADE) greatly increase the rate at which X-ray diffraction data can be acquired from crystals. One promising high throughput screening application of ADE is to rapidly combine protein crystals with fragment libraries. In this approach, each fragment soaks into a protein crystal either directly on data collection media or on a moving conveyor belt which then delivers the crystals to the X-ray beam. By simultaneously handling multiple crystals combined with fragment specimens, these techniques relax the automounter duty-cycle bottleneck that currently prevents optimal exploitation of third generation synchrotrons. Two factors limit the speed and scope of projects that are suitable for fragment screening using techniques such as ADE. Firstly, in applications where the high throughput screening apparatus is located inside the X-ray station (such as the conveyor belt system described above), the speed of data acquisition is limited by the time required for each fragment to soak into its protein crystal. Secondly, in applications where crystals are combined with fragments directly on data acquisition media (including both of the ADE methods described above), the maximum time that fragments have to soak into crystals is limited by evaporative dehydration of the protein crystals during the fragment soak. Here we demonstrate that both of these problems can be minimized by using small crystals, because the soak time required for a fragment hit to attain high occupancy depends approximately linearly on crystal size.
authors.affiliation: Office of Educational Programs, Brookhaven National Laboratory, Upton, New York, United States of America; Purchase College, State University of New York, Purchase, New York, United States of America.
authors.forename: Krystal
authors.initials: K
authors.lastname: Cole
authors.affiliation: Photon Sciences Directorate, Brookhaven National Laboratory, Upton, New York, United States of America.
authors.forename: Christian G
authors.initials: CG
authors.lastname: Roessler
authors.affiliation: Office of Educational Programs, Brookhaven National Laboratory, Upton, New York, United States of America; Freeport High School, Freeport, New York, United States of America.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Mulé
authors.affiliation: Office of Educational Programs, Brookhaven National Laboratory, Upton, New York, United States of America; Georgetown Day School, Washington, DC, United States of America.
authors.forename: Emma J
authors.initials: EJ
authors.lastname: Benson-Xu
authors.affiliation: Office of Educational Programs, Brookhaven National Laboratory, Upton, New York, United States of America; Physics Department, University of Oregon, Eugene, Oregon, United States of America.
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Mullen
authors.affiliation: Office of Educational Programs, Brookhaven National Laboratory, Upton, New York, United States of America; Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia, United States of America.
authors.forename: Benjamin A
authors.initials: BA
authors.lastname: Le
authors.affiliation: Office of Educational Programs, Brookhaven National Laboratory, Upton, New York, United States of America; Department of Biological Sciences, University of Delaware, Newark, Delaware, United States of America.
authors.forename: Alanna M
authors.initials: AM
authors.lastname: Tieman
authors.affiliation: Babylon Junior-Senior High School, Babylon, New York, United States of America.
authors.forename: Claire
authors.initials: C
authors.lastname: Birone
authors.affiliation: Sayville High School, West Sayville, New York, United States of America.
authors.forename: Maria
authors.initials: M
authors.lastname: Brown
authors.affiliation: Queens Metropolitan High School, Forest Hills, New York, United States of America.
authors.forename: Jesus
authors.initials: J
authors.lastname: Hernandez
authors.affiliation: Shoreham-Wading River High School, Shoreham, New York, United States of America.
authors.forename: Sherry
authors.initials: S
authors.lastname: Neff
authors.affiliation: Shelter Island High School, Shelter Island, New York, United States of America.
authors.forename: Daniel
authors.initials: D
authors.lastname: Williams
authors.affiliation: Photon Sciences Directorate, Brookhaven National Laboratory, Upton, New York, United States of America.
authors.forename: Marc
authors.initials: M
authors.lastname: Allaire
authors.affiliation: Photon Sciences Directorate, Brookhaven National Laboratory, Upton, New York, United States of America; Biosciences Department, Brookhaven National Laboratory, Upton, New York, United States of America.
authors.forename: Allen M
authors.initials: AM
authors.lastname: Orville
authors.affiliation: Photon Sciences Directorate, Brookhaven National Laboratory, Upton, New York, United States of America; Biosciences Department, Brookhaven National Laboratory, Upton, New York, United States of America.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Sweet
authors.affiliation: Photon Sciences Directorate, Brookhaven National Laboratory, Upton, New York, United States of America.
authors.forename: Alexei S
authors.initials: AS
authors.lastname: Soares
mesh_terms: D000162:Acoustics; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D011506:Proteins / Q000737:chemistry*
chemical_list: D011506:Proteins