Observation of arginyl-deoxyoligonucleotide interactions in TaqI endonuclease by detection of specific 1H NMR signals from 140kD [Nη1, Nη2, 15N Arg]TaqI/oligomer complexes

John Glushka, Francis Barany, David Cowburn

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Proton and nitrogen signals of the guanidinium amines in [Nη1, Nη2 15N Arg]TaqI endonuclease were observed using isotope filtered experiments and proton detected 1H{15N} heterocorrelated two dimensional NMR spectroscopy. These rapidly exchanging protons could be detected in the free enzyme only at pH 4.5; at pH 8.5, no signals were measured after extensive signal averaging. Addition of deoxyribonucleotide oligomers resulted in the appearance of two groups of signals at about 6.8 and 7.5 ppm. Since these signals are independent of the presence of cognate sequence or Mg2+, it is assumed they represent nonspecific arginyl-DNA interactions. This labeling/NMR approach provides a new method for investigating the role of arginine in protein-DNA interactions.

Original languageEnglish (US)
Pages (from-to)88-93
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume164
Issue number1
DOIs
StatePublished - Oct 16 1989
Externally publishedYes

Fingerprint

Oligomers
Protons
Nuclear magnetic resonance
Observation
Deoxyribonucleotides
DNA
Guanidine
Isotopes
Labeling
Nuclear magnetic resonance spectroscopy
Amines
Arginine
Magnetic Resonance Spectroscopy
Nitrogen
Enzymes
TCGA-specific type II deoxyribonucleases
Proton Magnetic Resonance Spectroscopy
Proteins
Experiments

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

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title = "Observation of arginyl-deoxyoligonucleotide interactions in TaqI endonuclease by detection of specific 1H NMR signals from 140kD [Nη1, Nη2, 15N Arg]TaqI/oligomer complexes",
abstract = "Proton and nitrogen signals of the guanidinium amines in [Nη1, Nη2 15N Arg]TaqI endonuclease were observed using isotope filtered experiments and proton detected 1H{15N} heterocorrelated two dimensional NMR spectroscopy. These rapidly exchanging protons could be detected in the free enzyme only at pH 4.5; at pH 8.5, no signals were measured after extensive signal averaging. Addition of deoxyribonucleotide oligomers resulted in the appearance of two groups of signals at about 6.8 and 7.5 ppm. Since these signals are independent of the presence of cognate sequence or Mg2+, it is assumed they represent nonspecific arginyl-DNA interactions. This labeling/NMR approach provides a new method for investigating the role of arginine in protein-DNA interactions.",
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T1 - Observation of arginyl-deoxyoligonucleotide interactions in TaqI endonuclease by detection of specific 1H NMR signals from 140kD [Nη1, Nη2, 15N Arg]TaqI/oligomer complexes

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AU - Barany, Francis

AU - Cowburn, David

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N2 - Proton and nitrogen signals of the guanidinium amines in [Nη1, Nη2 15N Arg]TaqI endonuclease were observed using isotope filtered experiments and proton detected 1H{15N} heterocorrelated two dimensional NMR spectroscopy. These rapidly exchanging protons could be detected in the free enzyme only at pH 4.5; at pH 8.5, no signals were measured after extensive signal averaging. Addition of deoxyribonucleotide oligomers resulted in the appearance of two groups of signals at about 6.8 and 7.5 ppm. Since these signals are independent of the presence of cognate sequence or Mg2+, it is assumed they represent nonspecific arginyl-DNA interactions. This labeling/NMR approach provides a new method for investigating the role of arginine in protein-DNA interactions.

AB - Proton and nitrogen signals of the guanidinium amines in [Nη1, Nη2 15N Arg]TaqI endonuclease were observed using isotope filtered experiments and proton detected 1H{15N} heterocorrelated two dimensional NMR spectroscopy. These rapidly exchanging protons could be detected in the free enzyme only at pH 4.5; at pH 8.5, no signals were measured after extensive signal averaging. Addition of deoxyribonucleotide oligomers resulted in the appearance of two groups of signals at about 6.8 and 7.5 ppm. Since these signals are independent of the presence of cognate sequence or Mg2+, it is assumed they represent nonspecific arginyl-DNA interactions. This labeling/NMR approach provides a new method for investigating the role of arginine in protein-DNA interactions.

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