C3 exoenzyme from Clostridium botulinum: structure of a tetragonal crystal form and a reassessment of NAD-induced flexure.
C3 exoenzyme from Clostridium botulinum (C3bot1) ADP-ribosylates and thereby inactivates Rho A, B and C GTPases in mammalian cells. The structure of a tetragonal crystal form has been determined by molecular replacement and refined to 1.89 A resolution. It is very similar to the apo structures determined previously from two different monoclinic crystal forms. An objective reassessment of available apo and nucleotide-bound C3bot1 structures indicates that, contrary to a previous report, the protein possesses a rigid core formed largely of beta-strands and that the general flexure that accompanies NAD binding is concentrated in two peripheral lobes. Tetragonal crystals disintegrate in the presence of NAD, most likely because of disruption of essential crystal contacts.
Department of Biology and Biochemistry, University of Bath, Claverton Down, BA2 7AY, England.
Hazel R
HR
Evans
Daniel E
DE
Holloway
J Mark
JM
Sutton
Joanne
J
Ayriss
Clifford C
CC
Shone
K Ravi
KR
Acharya
D036002:ADP Ribose Transferases / Q000737:chemistry* / Q000378:metabolism*; D001905:Botulinum Toxins / Q000737:chemistry* / Q000378:metabolism*; D003014:Clostridium botulinum / Q000201:enzymology*; D003460:Crystallization; D018360:Crystallography, X-Ray; D008024:Ligands; D008958:Models, Molecular; D009243:NAD / Q000378:metabolism*; D018583:Pliability; D011487:Protein Conformation
D008024:Ligands; D009243:NAD; D036002:ADP Ribose Transferases; C063007:exoenzyme C3, Clostridium botulinum; D001905:Botulinum Toxins