Spin-Labeled Analogs of CMP-NeuAc as NMR Probes of the α-2,6-Sialyltransferase ST6Gal I

Shan Liu, Andre Venot, Lu Meng, Fang Tian, Kelley W. Moremen, Geert Jan Boons, James H. Prestegard

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Structural data on mammalian proteins are often difficult to obtain by conventional NMR approaches because of an inability to produce samples with uniform isotope labeling in bacterial expression hosts. Proteins with sparse isotope labels can be produced in eukaryotic hosts by using isotope-labeled forms of specific amino acids, but structural analysis then requires information from experiments other than nuclear Overhauser effects. One source of alternate structural information is distance-dependent perturbation of spin relaxation times by nitroxide spin-labeled analogs of natural protein ligands. Here, we introduce spin-labeled analogs of sugar nucleotide donors for sialyltransferases, specifically, CMP-TEMPO (CMP-4-O-[2,2,6,6-tetramethylpiperidine-1-oxyl]) and CMP-4carboxyTEMPO (CMP-4-O-[4-carboxy-2,2,6,6-tetramethylpiperidinine-1-oxyl]). An ability to identify resonances from active site residues and produce distance constraints is illustrated on a 15N phenylalanine-labeled version of the structurally uncharacterized, α-2,6-linked sialyltransferase, ST6Gal I.

Original languageEnglish (US)
Pages (from-to)409-418
Number of pages10
JournalChemistry and Biology
Volume14
Issue number4
DOIs
StatePublished - Apr 20 2007

Fingerprint

Sialyltransferases
Cytidine Monophosphate
Isotopes
Nuclear magnetic resonance
Isotope Labeling
Proteins
Phenylalanine
Structural analysis
Sugars
Relaxation time
Labeling
Labels
Catalytic Domain
Nucleotides
Ligands
Amino Acids
beta-D-galactoside alpha 2-6-sialyltransferase
Experiments
TEMPO

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry

Cite this

Liu, Shan ; Venot, Andre ; Meng, Lu ; Tian, Fang ; Moremen, Kelley W. ; Boons, Geert Jan ; Prestegard, James H. / Spin-Labeled Analogs of CMP-NeuAc as NMR Probes of the α-2,6-Sialyltransferase ST6Gal I. In: Chemistry and Biology. 2007 ; Vol. 14, No. 4. pp. 409-418.
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Spin-Labeled Analogs of CMP-NeuAc as NMR Probes of the α-2,6-Sialyltransferase ST6Gal I. / Liu, Shan; Venot, Andre; Meng, Lu; Tian, Fang; Moremen, Kelley W.; Boons, Geert Jan; Prestegard, James H.

In: Chemistry and Biology, Vol. 14, No. 4, 20.04.2007, p. 409-418.

Research output: Contribution to journalArticle

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AU - Liu, Shan

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AU - Meng, Lu

AU - Tian, Fang

AU - Moremen, Kelley W.

AU - Boons, Geert Jan

AU - Prestegard, James H.

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