1nli: Difference between revisions

New page: left|200px<br /><applet load="1nli" size="450" color="white" frame="true" align="right" spinBox="true" caption="1nli, resolution 1.93Å" /> '''Complex of [E160A-E1...
 
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'''Complex of [E160A-E189A] trichosanthin and adenine'''<br />
'''Complex of [E160A-E189A] trichosanthin and adenine'''<br />


==Overview==
==Overview==
Trichosanthin is a ribosome-inactivating protein that cleaves specifically, the N-glycosidic bond of A-4324 of 28S rRNA. Trichosanthin and its variant, [E160A-E189A]-trichosanthin were found to bind an adenine base with a K(d), value of approximately 0.2mM. To determine how this doubly mutated variant, of trichosanthin interacts with adenine, the co-crystal structure of, [E160A-E189A]-trichosanthin and adenine was resolved to 0.193nm which, revealed that the active site conformation of the doubly mutated variant, is isomorphous to wild-type trichosanthin. Water molecules were found at, locations corresponding to the eliminated side chain of Glu-160 and, Glu-189. On the other hand, the adenine base interacted with, [E160A-E189A]-trichosanthin in a manner similar to that in wild-type, trichosanthin. Our structural analysis illustrates that Glu-160 and, Glu-189 in trichosanthin do not play an important role in maintaining the, active site conformation and binding adenine, an essential step for, substrate-enzyme interaction. On the other hand, removal of two glutamate, residues changed a large patch of negatively charged surface to a positive, charge, which may account for the destabilization of the oxocarbenium-like, transition-state and the significant decrease in ribosome-inactivating, activity in [E160A-E189A]-trichosanthin.
Trichosanthin is a ribosome-inactivating protein that cleaves specifically the N-glycosidic bond of A-4324 of 28S rRNA. Trichosanthin and its variant [E160A-E189A]-trichosanthin were found to bind an adenine base with a K(d) value of approximately 0.2mM. To determine how this doubly mutated variant of trichosanthin interacts with adenine, the co-crystal structure of [E160A-E189A]-trichosanthin and adenine was resolved to 0.193nm which revealed that the active site conformation of the doubly mutated variant is isomorphous to wild-type trichosanthin. Water molecules were found at locations corresponding to the eliminated side chain of Glu-160 and Glu-189. On the other hand, the adenine base interacted with [E160A-E189A]-trichosanthin in a manner similar to that in wild-type trichosanthin. Our structural analysis illustrates that Glu-160 and Glu-189 in trichosanthin do not play an important role in maintaining the active site conformation and binding adenine, an essential step for substrate-enzyme interaction. On the other hand, removal of two glutamate residues changed a large patch of negatively charged surface to a positive charge, which may account for the destabilization of the oxocarbenium-like transition-state and the significant decrease in ribosome-inactivating activity in [E160A-E189A]-trichosanthin.


==About this Structure==
==About this Structure==
1NLI is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Trichosanthes_kirilowii Trichosanthes kirilowii] with ADE as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/rRNA_N-glycosylase rRNA N-glycosylase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.2.22 3.2.2.22] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1NLI OCA].  
1NLI is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Trichosanthes_kirilowii Trichosanthes kirilowii] with <scene name='pdbligand=ADE:'>ADE</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/rRNA_N-glycosylase rRNA N-glycosylase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.2.22 3.2.2.22] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1NLI OCA].  


==Reference==
==Reference==
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[[Category: Trichosanthes kirilowii]]
[[Category: Trichosanthes kirilowii]]
[[Category: rRNA N-glycosylase]]
[[Category: rRNA N-glycosylase]]
[[Category: Chan, D.S.B.]]
[[Category: Chan, D S.B.]]
[[Category: Shaw, P.C.]]
[[Category: Shaw, P C.]]
[[Category: Williams, R.L.]]
[[Category: Williams, R L.]]
[[Category: Wong, K.B.]]
[[Category: Wong, K B.]]
[[Category: ADE]]
[[Category: ADE]]
[[Category: protein-dna complex]]
[[Category: protein-dna complex]]
[[Category: ribosome-inactivating protein]]
[[Category: ribosome-inactivating protein]]


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