A Phosphoenzyme Mimic, Overlapping Catalytic Sites and Reaction Coordinate Motion for Human NAMPT

Burgos, E; Ho, M; Almo, S; Schramm, V

doi:10.1073/pnas.0903898106

Title: A Phosphoenzyme Mimic, Overlapping Catalytic Sites and Reaction Coordinate Motion for Human NAMPT

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Abstract

Nicotinamide phosphoribosyltransferase (NAMPT) is highly evolved to capture nicotinamide (NAM) and replenish the nicotinamide adenine dinucleotide (NAD+) pool during ADP-ribosylation and transferase reactions. ATP-phosphorylation of an active-site histidine causes catalytic activation, increasing NAM affinity by 160,000. Crystal structures of NAMPT with catalytic site ligands identify the phosphorylation site, establish its role in catalysis, demonstrate unique overlapping ATP and phosphoribosyltransferase sites, and establish reaction coordinate motion. NAMPT structures with beryllium fluoride indicate a covalent H247-BeF3- as the phosphohistidine mimic. Activation of NAMPT by H247-phosphorylation causes stabilization of the enzyme-phosphoribosylpyrophosphate complex, permitting efficient capture of NAM. Reactant and product structures establish reaction coordinate motion for NAMPT to be migration of the ribosyl anomeric carbon from the pyrophosphate leaving group to the nicotinamide-N1 while the 5-phosphoryl group, the pyrophosphate moiety, and the nicotinamide ring remain fixed in the catalytic site.

Authors:: Burgos, E; Ho, M; Almo, S; Schramm, V

Publication Date:: Thu Jan 01 00:00:00 EST 2009

Research Org.:: Brookhaven National Lab. (BNL), Upton, NY (United States). National Synchrotron Light Source

Sponsoring Org.:: Doe - Office Of Science

OSTI Identifier:: 979985

Report Number(s):: BNL-92903-2010-JA
TRN: US201015%%1370

DOE Contract Number:: DE-AC02-98CH10886

Resource Type:: Journal Article

Journal Name:: Proceedings of the National Academy of Sciences of the United States of America

Additional Journal Information:: Journal Volume: 106; Journal Issue: 33

Country of Publication:: United States

Language:: English

Subject:: 36 MATERIALS SCIENCE; 59 BASIC BIOLOGICAL SCIENCES; 99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE; AFFINITY; ATP; BERYLLIUM FLUORIDES; CAPTURE; CARBON; CATALYSIS; COORDINATES; CRYSTAL STRUCTURE; HISTIDINE; HUMAN POPULATIONS; LIGANDS; MIGRATION; MOTION; NAD; NICOTINAMIDE; PHOSPHORYLATION; PONDS; PROGRAMMING LANGUAGES; PYROPHOSPHATES; RINGS; STABILIZATION; TRANSFERASES; national synchrotron light source

Citation Formats


                    Burgos, E, Ho, M, Almo, S, and Schramm, V. A Phosphoenzyme Mimic, Overlapping Catalytic Sites and Reaction Coordinate Motion for Human NAMPT.  United States: N. p., 2009. 
        Web.  doi:10.1073/pnas.0903898106.

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                    Burgos, E, Ho, M, Almo, S, & Schramm, V. A Phosphoenzyme Mimic, Overlapping Catalytic Sites and Reaction Coordinate Motion for Human NAMPT.  United States.  https://doi.org/10.1073/pnas.0903898106

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                    Burgos, E, Ho, M, Almo, S, and Schramm, V. 2009.  
        "A Phosphoenzyme Mimic, Overlapping Catalytic Sites and Reaction Coordinate Motion for Human NAMPT".  United States.  https://doi.org/10.1073/pnas.0903898106.

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@article{osti_979985,

  title        = {A Phosphoenzyme Mimic, Overlapping Catalytic Sites and Reaction Coordinate Motion for Human NAMPT},

  author       = {Burgos, E and Ho, M and Almo, S and Schramm, V},

  abstractNote = {Nicotinamide phosphoribosyltransferase (NAMPT) is highly evolved to capture nicotinamide (NAM) and replenish the nicotinamide adenine dinucleotide (NAD+) pool during ADP-ribosylation and transferase reactions. ATP-phosphorylation of an active-site histidine causes catalytic activation, increasing NAM affinity by 160,000. Crystal structures of NAMPT with catalytic site ligands identify the phosphorylation site, establish its role in catalysis, demonstrate unique overlapping ATP and phosphoribosyltransferase sites, and establish reaction coordinate motion. NAMPT structures with beryllium fluoride indicate a covalent H247-BeF3- as the phosphohistidine mimic. Activation of NAMPT by H247-phosphorylation causes stabilization of the enzyme-phosphoribosylpyrophosphate complex, permitting efficient capture of NAM. Reactant and product structures establish reaction coordinate motion for NAMPT to be migration of the ribosyl anomeric carbon from the pyrophosphate leaving group to the nicotinamide-N1 while the 5-phosphoryl group, the pyrophosphate moiety, and the nicotinamide ring remain fixed in the catalytic site.},

  doi          = {10.1073/pnas.0903898106},

  url          = {https://www.osti.gov/biblio/979985},
  journal      = {Proceedings of the National Academy of Sciences of the United States of America},
number       = 33,

  volume       = 106,

  place        = {United States},

  year         = {Thu Jan 01 00:00:00 EST 2009},

  month        = {Thu Jan 01 00:00:00 EST 2009}

}

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