1gzf
From Proteopedia
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==Overview== | ==Overview== | ||
| - | We have solved the crystal structures of Clostridium botulinum C3 | + | We have solved the crystal structures of Clostridium botulinum C3 exoenzyme free and complexed to NAD in the same crystal form, at 2.7 and 1.95 A, respectively. The asymmetric unit contains four molecules, which, in the free form, share the same conformation. Upon NAD binding, C3 underwent various conformational changes, whose amplitudes were differentially limited in the four molecules of the crystal unit. A major rearrangement concerns the loop that contains the functionally important ARTT motif (ADP-ribosyltransferase toxin turn-turn). The ARTT loop undergoes an ample swinging motion to adopt a conformation that covers the nicotinamide moiety of NAD. In particular, Gln-212, which belongs to the ARTT motif, flips over from a solvent-exposed environment to a buried conformation in the NAD binding pocket. Mutational experiments showed that Gln-212 is neither involved in NAD binding nor in the NAD-glycohydrolase activity of C3, whereas it plays a critical role in the ADP-ribosyl transfer to the substrate Rho. We observed additional NAD-induced movements, including a crab-claw motion of a subdomain that closes the NAD binding pocket. The data emphasized a remarkable NAD-induced plasticity of the C3 binding pocket and suggest that the NAD-induced ARTT loop conformation may be favored by the C3-NAD complex to bind to the substrate Rho. Our structural observations, together with a number of mutational experiments suggest that the mechanisms of Rho ADP-ribosylation by C3-NAD may be more complex than initially anticipated. |
==About this Structure== | ==About this Structure== | ||
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[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Boquet, P.]] | [[Category: Boquet, P.]] | ||
| - | [[Category: Charbonnier, J | + | [[Category: Charbonnier, J B.]] |
| - | [[Category: Du, M | + | [[Category: Du, M H.Le.]] |
[[Category: Flatau, G.]] | [[Category: Flatau, G.]] | ||
[[Category: Gas, F.]] | [[Category: Gas, F.]] | ||
[[Category: Menetrey, J.]] | [[Category: Menetrey, J.]] | ||
[[Category: Menez, A.]] | [[Category: Menez, A.]] | ||
| - | [[Category: Stura, E | + | [[Category: Stura, E A.]] |
| - | [[Category: Teulon, J | + | [[Category: Teulon, J M.]] |
[[Category: ADP]] | [[Category: ADP]] | ||
[[Category: NAD]] | [[Category: NAD]] | ||
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[[Category: c3 exoenzyme]] | [[Category: c3 exoenzyme]] | ||
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 12:55:40 2008'' |
Revision as of 10:55, 21 February 2008
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STRUCTURE OF THE CLOSTRIDIUM BOTULINUM C3 EXOENZYME (WILD-TYPE) IN COMPLEX WITH NAD
Overview
We have solved the crystal structures of Clostridium botulinum C3 exoenzyme free and complexed to NAD in the same crystal form, at 2.7 and 1.95 A, respectively. The asymmetric unit contains four molecules, which, in the free form, share the same conformation. Upon NAD binding, C3 underwent various conformational changes, whose amplitudes were differentially limited in the four molecules of the crystal unit. A major rearrangement concerns the loop that contains the functionally important ARTT motif (ADP-ribosyltransferase toxin turn-turn). The ARTT loop undergoes an ample swinging motion to adopt a conformation that covers the nicotinamide moiety of NAD. In particular, Gln-212, which belongs to the ARTT motif, flips over from a solvent-exposed environment to a buried conformation in the NAD binding pocket. Mutational experiments showed that Gln-212 is neither involved in NAD binding nor in the NAD-glycohydrolase activity of C3, whereas it plays a critical role in the ADP-ribosyl transfer to the substrate Rho. We observed additional NAD-induced movements, including a crab-claw motion of a subdomain that closes the NAD binding pocket. The data emphasized a remarkable NAD-induced plasticity of the C3 binding pocket and suggest that the NAD-induced ARTT loop conformation may be favored by the C3-NAD complex to bind to the substrate Rho. Our structural observations, together with a number of mutational experiments suggest that the mechanisms of Rho ADP-ribosylation by C3-NAD may be more complex than initially anticipated.
About this Structure
1GZF is a Single protein structure of sequence from Clostridium botulinum with , , and as ligands. Known structural/functional Site: . Full crystallographic information is available from OCA.
Reference
NAD binding induces conformational changes in Rho ADP-ribosylating clostridium botulinum C3 exoenzyme., Menetrey J, Flatau G, Stura EA, Charbonnier JB, Gas F, Teulon JM, Le Du MH, Boquet P, Menez A, J Biol Chem. 2002 Aug 23;277(34):30950-7. Epub 2002 May 23. PMID:12029083
Page seeded by OCA on Thu Feb 21 12:55:40 2008
Categories: Clostridium botulinum | Single protein | Boquet, P. | Charbonnier, J B. | Du, M H.Le. | Flatau, G. | Gas, F. | Menetrey, J. | Menez, A. | Stura, E A. | Teulon, J M. | ADP | NAD | NIR | SO4 | Adp-ribosyltransferase | Bacterial toxin | C3 exoenzyme
