Backbone 1H, 13C, 15N NMR assignments of yeast OMP synthase in unliganded form and in complex with orotidine 5'-monophosphate

Michael Riis Hansen, Richard Harris, Eric W. Barr, Hong Cheng, Mark E. Girvin, Charles Grubmeyer

Research output: Contribution to journalArticle

Abstract

The type I phosphoribosyltransferase OMP synthase (EC 2.4.2.10) is involved in de novo synthesis of pyrimidine nucleotides forming the UMP precursor orotidine 5'-monophosphate (OMP). The homodimeric enzyme has a Rossman α/β core topped by a base-enclosing "hood" domain and a flexible domain-swapped catalytic loop. High-resolution X-ray structures of the homologous Salmonella typhimurium and yeast enzymes show that a general compacting of the core as well as movement of the hood and a major disorder-to-order transition of the loop occur upon binding of ligands MgPRPP and orotate. Here we present backbone NMR assignments for the unliganded yeast enzyme (49 kDa) and its complex with product OMP. We were able to assign 212-213 of the 225 non-proline backbone 15N and amide proton resonances. Significant difference in chemical shifts of the amide cross peaks occur in regions of the structure that undergo movement upon ligand occupancy in the S. typhimurium enzyme.

Original languageEnglish (US)
Pages (from-to)103-108
Number of pages6
JournalBiomolecular NMR Assignments
Volume8
Issue number1
DOIs
StatePublished - 2014

Fingerprint

Yeast
Yeasts
Nuclear magnetic resonance
Enzymes
Salmonella typhimurium
Amides
Orotate Phosphoribosyltransferase
Pyrimidine Nucleotides
Uridine Monophosphate
Ligands
Salmonella
Chemical shift
Protons
Catalytic Domain
X-Rays
X rays
orotidylic acid
Carbon-13 Magnetic Resonance Spectroscopy

Keywords

  • Domain swapping
  • Enzyme conformational dynamics
  • NMR resonance assignments
  • OMP synthase
  • Phosphoribosyltransferase

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry

Cite this

Backbone 1H, 13C, 15N NMR assignments of yeast OMP synthase in unliganded form and in complex with orotidine 5'-monophosphate. / Hansen, Michael Riis; Harris, Richard; Barr, Eric W.; Cheng, Hong; Girvin, Mark E.; Grubmeyer, Charles.

In: Biomolecular NMR Assignments, Vol. 8, No. 1, 2014, p. 103-108.

Research output: Contribution to journalArticle

Hansen, Michael Riis ; Harris, Richard ; Barr, Eric W. ; Cheng, Hong ; Girvin, Mark E. ; Grubmeyer, Charles. / Backbone 1H, 13C, 15N NMR assignments of yeast OMP synthase in unliganded form and in complex with orotidine 5'-monophosphate. In: Biomolecular NMR Assignments. 2014 ; Vol. 8, No. 1. pp. 103-108.
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