X-ray structure determination of the glycine cleavage system protein H of Mycobacterium tuberculosis using an inverse Compton synchrotron X-ray source.
Structural genomics discovery projects require ready access to both X-ray diffraction and NMR spectroscopy which support the collection of experimental data needed to solve large numbers of novel protein structures. The most productive X-ray crystal structure determination laboratories make extensive use of tunable synchrotron X-ray light to solve novel structures by anomalous diffraction methods. This requires that frozen cryo-protected crystals be shipped to large multi acre synchrotron facilities for data collection. In this paper we report on the development and use of the first laboratory-scale synchrotron light source capable of performing many of the state-of-the-art synchrotron applications in X-ray science. This Compact Light Source is a first-in-class device that uses inverse Compton scattering to generate X-rays of sufficient flux, tunable wavelength and beam size to allow high-resolution X-ray diffraction data collection from protein crystals. We report on benchmarking tests of X-ray diffraction data collection with hen egg white lysozyme, and the successful high-resolution X-ray structure determination of the Glycine cleavage system protein H from Mycobacterium tuberculosis using diffraction data collected with the Compact Light Source X-ray beam.
Abendroth
Jan
J
Emerald BioStructures, 7869 NE Day Road West, Bainbridge Island, WA 98110, USA.
McCormick
Michael S
MS
Edwards
Thomas E
TE
Staker
Bart
B
Loewen
Roderick
R
Gifford
Martin
M
Rifkin
Jeff
J
Mayer
Chad
C
Guo
Wenjin
W
Zhang
Yang
Y
Myler
Peter
P
Kelley
Angela
A
Analau
Erwin
E
Hewitt
Stephen Nakazawa
SN
Napuli
Alberto J
AJ
Kuhn
Peter
P
Ruth
Ronald D
RD
Stewart
Lance J
LJ
D000594:Amino Acid Oxidoreductases; D002352:Carrier Proteins; D050961:Glycine Decarboxylase Complex H-Protein; D009097:Multienzyme Complexes; D009113:Muramidase; D009169:Mycobacterium tuberculosis / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D017356:Synchrotrons; D014166:Transferases; D014961:X-Ray Diffraction / Q000295:instrumentation / Q000379:methods*; D014965:X-Rays
D002352:Carrier Proteins; D050961:Glycine Decarboxylase Complex H-Protein; D009097:Multienzyme Complexes; D011506:Proteins; C064525:glycine cleavage system; D000594:Amino Acid Oxidoreductases; D014166:Transferases; C421864:hen egg lysozyme; D009113:Muramidase