Computational Design of an α-gliadin Peptidase

TitleComputational Design of an α-gliadin Peptidase
Publication TypeJournal Article
Year of Publication2012
AuthorsGordon, S. R., Stanley E. J., Wolf S., Toland A., Wu S. J., Hadidi D., Mills J. H., Baker D., Pultz I. S., & Siegel J. B.
JournalJournal of the American Chemical Society
Volume134
Issue50
Pagination20513-20
Date Published2012 Dec 19
ISSN1520-5126
KeywordsAmino Acid Sequence, Gliadin, Models, Molecular, Molecular Sequence Data, Peptide Hydrolases, Primary Publication
Abstract

The ability to rationally modify enzymes to perform novel chemical transformations is essential for the rapid production of next-generation protein therapeutics. Here we describe the use of chemical principles to identify a naturally occurring acid-active peptidase, and the subsequent use of computational protein design tools to reengineer its specificity toward immunogenic elements found in gluten that are the proposed cause of celiac disease. The engineered enzyme exhibits a k(cat)/K(M) of 568 M(-1) s(-1), representing a 116-fold greater proteolytic activity for a model gluten tetrapeptide than the native template enzyme, as well as an over 800-fold switch in substrate specificity toward immunogenic portions of gluten peptides. The computationally engineered enzyme is resistant to proteolysis by digestive proteases and degrades over 95% of an immunogenic peptide implicated in celiac disease in under an hour. Thus, through identification of a natural enzyme with the pre-existing qualities relevant to an ultimate goal and redefinition of its substrate specificity using computational modeling, we were able to generate an enzyme with potential as a therapeutic for celiac disease.

URLhttp://www.ncbi.nlm.nih.gov/pubmed/23153249
DOI10.1021/ja3094795
Custom1

http://www.ncbi.nlm.nih.gov/pubmed/23153249?dopt=Abstract

Alternate JournalJ. Am. Chem. Soc.
Full Text

The ability to rationally modify enzymes to perform novel chemical transformations is essential for the rapid production of next-generation protein therapeutics. Here we describe the use of chemical principles to identify a naturally occurring acid-active peptidase, and the subsequent use of computational protein design tools to reengineer its specificity toward immunogenic elements found in gluten that are the proposed cause of celiac disease. The engineered enzyme exhibits a k(cat)/K(M) of 568 M(-1) s(-1), representing a 116-fold greater proteolytic activity for a model gluten tetrapeptide than the native template enzyme, as well as an over 800-fold switch in substrate specificity toward immunogenic portions of gluten peptides. The computationally engineered enzyme is resistant to proteolysis by digestive proteases and degrades over 95% of an immunogenic peptide implicated in celiac disease in under an hour. Thus, through identification of a natural enzyme with the pre-existing qualities relevant to an ultimate goal and redefinition of its substrate specificity using computational modeling, we were able to generate an enzyme with potential as a therapeutic for celiac disease.

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