The structure of the pantothenate kinase·ADP·pantothenate ternary complex reveals the relationship between the binding sites for substrate, allosteric regulator, and antimetabolites

Robert A. Ivey, Yong Mei Zhang, Kristopher G. Virga, Kirk Hevener, Richard E. Lee, Charles O. Rock, Suzanne Jackowski, Hee Won Park

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Pantothenate kinase catalyzes the first step in the biosynthesis of coenzyme A, the major acyl group carrier in biology. In bacteria, regulation of pantothenate kinase activity is a major factor in controlling intracellular coenzyme A levels, and pantothenate analogs are growth-inhibiting antimetabolites. We have extended the structural information on Escherichia coli pantothenate kinase by determining the structure of the enzyme· ADP· pantothenate ternary complex. Pantothenate binding induces a significant conformational change in amino acids 243-263, which form a "lid" that folds over the open pantothenate binding groove. The positioning of the substrates suggests the reaction proceeds by a concerted mechanism that involves a dissociative transition state, although the negative charge neutralization of the γ-phosphate by Arg-243, Lys-101, and Mg 2+ coupled with hydrogen bonding of the C1 of pantothenate to Asp-127 suggests different interpretations of the phosphoryl transfer mechanism of pantothenate kinase. N-alkylpantothenamides are substrates for pantothenate kinase. Modeling these antimetabolites into the pantothenate active site predicts that they bind in the same orientation as pantothenate with their alkyl chains interacting with the hydrophobic dome over the pantothenate pocket, which is also accessed by the 13-mercaptoethylamine moiety of the allosteric regulator, coenzyme A. These structural/biochemical studies illustrate the intimate relationship between the substrate, allosteric regulator, and antimetabolite binding sites on pantothenate kinase and provide a framework for studies of its catalysis and feedback regulation.

Original languageEnglish (US)
Pages (from-to)35622-35629
Number of pages8
JournalJournal of Biological Chemistry
Volume279
Issue number34
DOIs
StatePublished - Aug 20 2004

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Antimetabolites
Binding Sites
Substrates
Coenzyme A
Cysteamine
Biosynthesis
Domes
Hydrogen Bonding
Catalysis
Adenosine Diphosphate
Escherichia coli
pantothenate kinase
Catalytic Domain
Bacteria
Hydrogen bonds
Phosphates
Feedback
Amino Acids
Enzymes
Growth

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

The structure of the pantothenate kinase·ADP·pantothenate ternary complex reveals the relationship between the binding sites for substrate, allosteric regulator, and antimetabolites. / Ivey, Robert A.; Zhang, Yong Mei; Virga, Kristopher G.; Hevener, Kirk; Lee, Richard E.; Rock, Charles O.; Jackowski, Suzanne; Park, Hee Won.

In: Journal of Biological Chemistry, Vol. 279, No. 34, 20.08.2004, p. 35622-35629.

Research output: Contribution to journalArticle

Ivey, Robert A. ; Zhang, Yong Mei ; Virga, Kristopher G. ; Hevener, Kirk ; Lee, Richard E. ; Rock, Charles O. ; Jackowski, Suzanne ; Park, Hee Won. / The structure of the pantothenate kinase·ADP·pantothenate ternary complex reveals the relationship between the binding sites for substrate, allosteric regulator, and antimetabolites. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 34. pp. 35622-35629.
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