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| | {{STRUCTURE_1vpo| PDB=1vpo | SCENE= }} | | {{STRUCTURE_1vpo| PDB=1vpo | SCENE= }} |
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| - | '''Crystal Structure Analysis of the Anti-testosterone Fab in Complex with Testosterone'''
| + | ===Crystal Structure Analysis of the Anti-testosterone Fab in Complex with Testosterone=== |
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| - | ==Overview==
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| - | A highly selective, high affinity recombinant anti-testosterone Fab fragment has been generated by stepwise optimization of the complementarity-determining regions (CDRs) by random mutagenesis and phage display selection of a monoclonal antibody (3-C(4)F(5)). The best mutant (77 Fab) was obtained by evaluating the additivity effects of different independently selected CDR mutations. The 77 Fab contains 20 mutations and has about 40-fold increased affinity (K(d) = 3 x 10(-10) m) when compared with the wild-type (3-C(4)F(5)) Fab. To obtain structural insight into factors, which are needed to improve binding properties, we have determined the crystal structures of the mutant 77 Fab fragment with (2.15 A) and without testosterone (2.10 A) and compared these with previously determined wild-type structures. The overall testosterone binding of the 77 Fab is similar to that of the wild-type. The improved affinity and specificity of the 77 Fab fragment are due to more comprehensive packing of the testosterone with the protein, which is the result of small structural changes within the variable domains. Only one important binding site residue Glu-95 of the heavy chain CDR3 is mutated to alanine in the 77 Fab fragment. This mutation, originally selected from the phage library based on improved specificity, provides more free space for the testosterone D-ring. The light chain CDR1 of 77 Fab containing eight mutations has the most significant effect on the improved affinity, although it has no direct contact with the testosterone. The mutations of CDR-L1 cause a rearrangement in its conformation, leading to an overall fine reshaping of the binding site.
| + | The line below this paragraph, {{ABSTRACT_PUBMED_12196551}}, adds the Publication Abstract to the page |
| | + | (as it appears on PubMed at http://www.pubmed.gov), where 12196551 is the PubMed ID number. |
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| | + | {{ABSTRACT_PUBMED_12196551}} |
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| | ==About this Structure== | | ==About this Structure== |
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| | [[Category: Fab fragment]] | | [[Category: Fab fragment]] |
| | [[Category: Testosterone complex]] | | [[Category: Testosterone complex]] |
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 12:46:06 2008'' | + | |
| | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Mon Jul 28 12:25:41 2008'' |
Revision as of 09:25, 28 July 2008
Template:STRUCTURE 1vpo
Crystal Structure Analysis of the Anti-testosterone Fab in Complex with Testosterone
Template:ABSTRACT PUBMED 12196551
About this Structure
This structure supersedes the now removed PDB entry 1l7s. Full crystallographic information is available from OCA.
Reference
Crystal structure of an in vitro affinity- and specificity-matured anti-testosterone Fab in complex with testosterone. Improved affinity results from small structural changes within the variable domains., Valjakka J, Hemminki A, Niemi S, Soderlund H, Takkinen K, Rouvinen J, J Biol Chem. 2002 Nov 15;277(46):44021-7. Epub 2002 Aug 23. PMID:12196551
Page seeded by OCA on Mon Jul 28 12:25:41 2008
