Structure of glycosomal glyceraldehyde-3-phosphate dehydrogenase from Trypanosoma brucei determined from Laue data.
The three-dimensional structure of glycosomal glyceraldehyde-3-phosphate dehydrogenase [D-glyceraldehyde-3-phosphate:NAD+ oxidoreductase (phosphorylating), EC 1.12.1.12] from the sleeping-sickness parasite Trypanosoma brucei was solved by molecular replacement at 3.2-A resolution with an x-ray data set collected by the Laue method. For data collection, three crystals were exposed to the polychromatic synchrotron x-ray beam for a total of 20.5 sec. The structure was solved by using the Bacillus stearothermophilus enzyme model [Skarzyński, T., Moody, P. C. E. & Wonacott, A. J. (1987) J. Mol. Biol. 193, 171-187] with a partial data set which was 37% complete. The crystals contain six subunits per asymmetric unit, which allowed us to overcome the absence of > 60% of the reflections by 6-fold density averaging. After molecular dynamics refinement, the current molecular model has an R factor of 17.6%. Comparing the structure of the trypanosome enzyme with that of the homologous human muscle enzyme, which was determined at 2.4-A resolution, reveals important structural differences in the NAD binding region. These are of great interest for the design of specific inhibitors of the parasite enzyme.
Vellieux
F M
FM
Department of Chemistry, BIOSON Research Institute, University of Groningen, The Netherlands.
Hajdu
J
J
Verlinde
C L
CL
Groendijk
H
H
Read
R J
RJ
Greenhough
T J
TJ
Campbell
J W
JW
Kalk
K H
KH
Littlechild
J A
JA
Watson
H C
HC
D000818:Animals; D001665:Binding Sites; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D009243:NAD / Q000378:metabolism; D015388:Organelles / Q000201:enzymology*; D011487:Protein Conformation; D014346:Trypanosoma brucei brucei / Q000201:enzymology*; D014961:X-Ray Diffraction / Q000379:methods
D009243:NAD; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases