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NMR detection of protein 15 N spins near paramagnetic lanthanide ions

John, Michael; Park, Ah Young; Dixon, Nicholas; Otting, Gottfried

Description

Pronounced paramagnetic relaxation enhancement (PRE) due to paramagnetic metal ions prevents the observation of NMR signals from 1H spins near the metal. While 15N spins are less prone to PRE, the intrinsic sensitivity of 15N NMR spectroscopy is low. This Communication presents a 1H detected out-and-back Nz-exchange experiment which allows the measurement of pseudocontact shifts of 15N spins located as close as 6 Å from a Dy3+ ion in a 30 kDa protein complex. The experiment relies on the...[Show more]

dc.contributor.authorJohn, Michael
dc.contributor.authorPark, Ah Young
dc.contributor.authorDixon, Nicholas
dc.contributor.authorOtting, Gottfried
dc.date.accessioned2015-12-07T22:44:52Z
dc.identifier.issn0002-7863
dc.identifier.urihttp://hdl.handle.net/1885/25382
dc.description.abstractPronounced paramagnetic relaxation enhancement (PRE) due to paramagnetic metal ions prevents the observation of NMR signals from 1H spins near the metal. While 15N spins are less prone to PRE, the intrinsic sensitivity of 15N NMR spectroscopy is low. This Communication presents a 1H detected out-and-back Nz-exchange experiment which allows the measurement of pseudocontact shifts of 15N spins located as close as 6 Å from a Dy3+ ion in a 30 kDa protein complex. The experiment relies on the chemical exchange between paramagnetic and diamagnetic metal ions during two mixing times during which the 15N magnetization is stored as PRE-insensitive longitudinal magnetization. It is demonstrated with the complex between the subunit θ and the N-terminal domain of the subunit ε of E. coli DNA polymerase III, prepared with a mixture of Dy3+ and La3+. Pseudocontact shifts were measured for 61 15N spins which were not observable in a conventional 15N-HSQC spectrum.
dc.publisherAmerican Chemical Society
dc.sourceJournal of the American Chemical Society
dc.subjectKeywords: DNA polymerase; lanthanide; amino terminal sequence; anisotropy; article; complex formation; dissociation constant; magnetic field; nitrogen nuclear magnetic resonance; protein analysis; protein domain; proton nuclear magnetic resonance; Ions; Lanthanoid
dc.titleNMR detection of protein 15 N spins near paramagnetic lanthanide ions
dc.typeJournal article
local.description.notesImported from ARIES
local.identifier.citationvolume129
dc.date.issued2007
local.identifier.absfor069999 - Biological Sciences not elsewhere classified
local.identifier.ariespublicationu4005981xPUB38
local.type.statusPublished Version
local.contributor.affiliationJohn, Michael, College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationPark, Ah Young, College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationDixon, Nicholas, College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationOtting, Gottfried, College of Physical and Mathematical Sciences, ANU
local.description.embargo2037-12-31
local.bibliographicCitation.issue3
local.bibliographicCitation.startpage462
local.bibliographicCitation.lastpage463
local.identifier.doi10.1021/ja066995o
dc.date.updated2015-12-07T11:29:36Z
local.identifier.scopusID2-s2.0-33846424285
CollectionsANU Research Publications

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