1nj9
From Proteopedia
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[[Image:1nj9.gif|left|200px]] | [[Image:1nj9.gif|left|200px]] | ||
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'''Cocaine hydrolytic antibody 15A10''' | '''Cocaine hydrolytic antibody 15A10''' | ||
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==About this Structure== | ==About this Structure== | ||
| - | + | Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1NJ9 OCA]. | |
==Reference== | ==Reference== | ||
Crystallographic and biochemical analysis of cocaine-degrading antibody 15A10., Larsen NA, de Prada P, Deng SX, Mittal A, Braskett M, Zhu X, Wilson IA, Landry DW, Biochemistry. 2004 Jun 29;43(25):8067-76. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/15209502 15209502] | Crystallographic and biochemical analysis of cocaine-degrading antibody 15A10., Larsen NA, de Prada P, Deng SX, Mittal A, Braskett M, Zhu X, Wilson IA, Landry DW, Biochemistry. 2004 Jun 29;43(25):8067-76. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/15209502 15209502] | ||
| - | [[Category: Mus musculus]] | ||
| - | [[Category: Protein complex]] | ||
[[Category: Deng, S X.]] | [[Category: Deng, S X.]] | ||
[[Category: Landry, D W.]] | [[Category: Landry, D W.]] | ||
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[[Category: Wilson, I A.]] | [[Category: Wilson, I A.]] | ||
[[Category: Zhu, X.]] | [[Category: Zhu, X.]] | ||
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| - | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 02:36:02 2008'' | |
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Revision as of 23:36, 2 May 2008
Cocaine hydrolytic antibody 15A10
Overview
Catalytic antibody 15A10 hydrolyzes the benzoyl ester of cocaine to form the nonpsychoactive metabolites benzoic acid and ecgonine methylester. Here, we report biochemical and structural studies that characterize the catalytic mechanism. The crystal structure of the cocaine-hydrolyzing monoclonal antibody (mAb) 15A10 has been determined at 2.35 A resolution. The binding pocket is fairly shallow and mainly hydrophobic but with a cluster of three hydrogen-bond donating residues (TrpL96, AsnH33, and TyrH35). Computational docking of the transition state analogue (TSA) indicates that these residues are appropriately positioned to coordinate the phosphonate moiety of the TSA and, hence, form an oxyanion hole. Tyrosine modification of the antibody with tetranitromethane reduced hydrolytic activity to background level. The contribution from these and other residues to catalysis and TSA binding was explored by site-directed mutagenesis of 15A10 expressed in a single chain fragment variable (scFv) format. The TyrH35Phe mutant had 4-fold reduced activity, and TrpL96Ala, TrpL96His, and AsnH33Ala mutants were all inactive. Comparison with an esterolytic antibody D2.3 revealed a similar arrangement of tryptophan, asparagine, and tyrosine residues in the oxyanion hole that stabilizes the transition state for ester hydrolysis. Furthermore, the crystal structure of the bacterial cocaine esterase (cocE) also showed that the cocE employs a tyrosine hydroxyl in the oxyanion hole. Thus, the biochemical and structural data are consistent with the catalytic antibody providing oxyanion stabilization as its major contribution to catalysis.
About this Structure
Full crystallographic information is available from OCA.
Reference
Crystallographic and biochemical analysis of cocaine-degrading antibody 15A10., Larsen NA, de Prada P, Deng SX, Mittal A, Braskett M, Zhu X, Wilson IA, Landry DW, Biochemistry. 2004 Jun 29;43(25):8067-76. PMID:15209502 Page seeded by OCA on Sat May 3 02:36:02 2008
