2ddf

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spinqualitylabelsall models 2ddf, resolution 1.700Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image

Crystal structure of TACE in complex with TAPI-2

Overview

The crystallization of TNF-alpha converting enzyme (TACE) has been useful, in understanding the structure-activity relationships of new chemical, entities. However, the propensity of TACE to undergo autoproteolysis has, made enzyme handling difficult and impeded the identification of inhibitor, soakable crystal forms. The autoproteolysis of TACE was found to be, specific (Y352-V353) and occurred within a flexible loop that is in close, proximity to the P-side of the active site. The rate of autoproteolysis, was found to be proportional to the concentration of TACE, suggesting a, bimolecular reaction mechanism. A limited specificity study of the S(1)', subsite was conducted using surrogate peptides and suggested substitutions, that would stabilize the proteolysis of the loop at positions Y352-V353., Two mutant proteases (V353G and V353S) were generated and proved to be, highly resistant to autoproteolysis. The kinetics of the more resistant, mutant (V353G) and wild-type TACE were compared and demonstrated virtually, identical IC(50) values for a panel of competitive inhibitors. However, the k(cat)/K(m) of the mutant for a larger substrate (P6 - P(6)') was, approximately 5-fold lower than that for the wild-type enzyme. Comparison, of the complexed wild-type and mutant structures indicated a subtle shift, in a peripheral P-side loop (comprising the mutation site) that may be, involved in substrate binding/turnover and might explain the mild kinetic, difference. The characterization of this stabilized form of TACE has, yielded an enzyme with similar native kinetic properties and identified a, novel crystal form that is suitable for inhibitor soaking and structure, determination.

About this Structure

2DDF is a Single protein structure of sequence from Homo sapiens with ZN, CA, INN, IMD, CIT and IPA as ligands. Active as ADAM 17 endopeptidase, with EC number 3.4.24.86 Full crystallographic information is available from OCA.

Reference

Stabilization of the autoproteolysis of TNF-alpha converting enzyme (TACE) results in a novel crystal form suitable for structure-based drug design studies., Ingram RN, Orth P, Strickland CL, Le HV, Madison V, Beyer BM, Protein Eng Des Sel. 2006 Apr;19(4):155-61. Epub 2006 Feb 3. PMID:16459338

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