Selective stabilization of the chorismate mutase transition state by a positively charged hydrogen bond donor.

Selective stabilization of the chorismate mutase transition state by a positively charged hydrogen bond donor. Research Abstract Details 

Research Abstract Table of Contents

Jump to the:

Selective stabilization of the chorismate mutase transition state by a positively charged hydrogen bond donor. Abstract Text:

Alexander ,Peter Kast,Donald Hilvert,

Citrulline was incorporated via chemical semisynthesis at position 90 in the active site of the AroH chorismate mutase from Bacillus subtilis. The wild-type arginine at this position makes hydrogen-bonding interactions with the ether oxygen of chorismate. Replacement of the positively charged guanidinium group with the isosteric but neutral urea has a dramatic effect on the ability of the enzyme to convert chorismate into prephenate. The Arg90Cit variant exhibits a >104-fold decrease in the catalytic rate constant kcat with a 2.7-fold increase in the Michaelis constant Km. In contrast, its affinity for a conformationally constrained inhibitor molecule that effectively mimics the geometry but not the dissociative character of the transition state is only reduced by a factor of approximately 6. These results show that an active site merely complementary to the reactive conformation of chorismate is insufficient for catalysis of the mutase reaction. Instead, electrostatic stabilization of the polarized transition state by provision of a cationic hydrogen bond donor proximal to the oxygen in the breaking C-O bond is essential for high catalytic efficiency.

Selective stabilization of the chorismate mutase transition state by a positively charged hydrogen bond donor. Publishing Authors By Initials

A ,P Kast,D Hilvert,

For similar proteins: recombinant proteins: recombinant fusion proteins research abstracts see: proteins: recombinant proteins: recombinant fusion proteins research

PUBMED ID PMID:

MEDLINE DATE:

Selective stabilization of the chorismate mutase transition state by a positively charged hydrogen bond donor. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Journal of the American Chemical Society

VOLUME: 125

Page Numbers: 3206-7

Journal Abbreviation: J. Am. Chem. Soc.

ISSN: 0002-7863

DAY: 19

MONTH: Mar

YEAR: 2003

Selective stabilization of the chorismate mutase transition state by a positively charged hydrogen bond donor. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 7503056

Selective stabilization of the chorismate mutase transition state by a positively charged hydrogen bond donor. Keywords Mesh Terms:

KEYWORDS: Recombinant Fusion Proteins

MESH TERMS: metabolism

Chemical & Substance for Abstract: Selective stabilization of the chorismate mutase transition state by a positively charged hydrogen bond donor. Information

Substance Name: Chorismate Mutase

Registry Number: EC 5.4.99.5

Grant and Affiliation Information for Selective stabilization of the chorismate mutase transition state by a positively charged hydrogen bond donor.

AFFILIATION: Laboratorium für Organische Chemie, Swiss Federal Institute of Technology, ETH Hönggerberg, CH-8093 Zürich, Switzerland.

Country: United States

AGENCY:

GRANT:

ACRONYM:

MEDLINETA: J Am Chem Soc

REFSOURCE:

DATABASENAME:

ACCESSION NUMBER:

Number Hits: 0

Selective stabilization of the chorismate mutase transition state by a positively charged hydrogen bond donor Related Publications

• A simple selection strategy for evolving highly efficient enzymes.
• A simple tagging system for protein encapsulation.
• Bifunctional catalysis of proton transfer at an antibody active site.
• Catalysis of oxidative protein folding by small-molecule diselenides.
• Charge optimization increases the potency and selectivity of a chorismate mutase inhibitor.
• Conversion of a PLP-dependent racemase into an aldolase by a single active site mutation.
• Isochorismate pyruvate lyase: a pericyclic reaction mechanism?
• Kinetics and thermodynamics of ligand binding to a molten globular enzyme and its native counterpart.
• Minimalist active-site redesign: teaching old enzymes new tricks.
• Novel enzymes through design and evolution.
• Protein design by directed evolution.
• Reinvestigation of a selenopeptide with purportedly high glutathione peroxidase activity.
• Selective stabilization of the chorismate mutase transition state by a positively charged hydrogen bond donor.
• Selenoglutaredoxin as a glutathione peroxidase mimic.
• Selenoglutathione: efficient oxidative protein folding by a diselenide.
• Structure and dynamics of a molten globular enzyme.
• Switching Antibody Specificity through Minimal Mutation.
• The putative Diels-Alderase macrophomate synthase is an efficient aldolase.