Applications of a slow-scan CCD camera in protein electron crystallography.
A Gatan 1024 x 1024 slow-scan charge-coupled device (CCD) camera has been interfaced to a JEOL4000EX electron cryomicroscope and explored for its usefulness in the electron crystallographic analysis of thin, glucose-embedded crystals of crotoxin complex kept at -125 degrees C. We show that the camera allows for an on-line assessment of the crystals' crystallinity, flatness, and thickness. Intensities obtained from electron diffraction patterns acquired with the camera have been statistically analyzed and were found to be consistent with theoretically expected values. A quantitative analysis of the diffraction intensity as function of the accumulated electron dose suggests the possibility of recording up to 250 diffraction patterns with 3.5 A resolution from a single crotoxin complex crystal 128 A thick. Tilt series of 125 electron diffraction patterns with 3.5 A data acquired from a single crystal are shown to be practically feasible. The current study demonstrates for the first time the effectiveness of using a slow-scan CCD camera for electron diffraction data collection from thin protein crystals at near atomic resolution.
Brink
J
J
Verna and Marrs McLean Department of Biochemistry, Baylor College of Medicine, Houston, Texas 77030.
Chiu
W
W
D003439:Crotoxin / Q000737:chemistry*; D003461:Crystallography; D008854:Microscopy, Electron / Q000295:instrumentation* / Q000379:methods; D010781:Photography / Q000295:instrumentation* / Q000379:methods; D011487:Protein Conformation; D011506:Proteins / Q000648:ultrastructure*
D011506:Proteins; D003439:Crotoxin