Direct NMR resonance assignments of the active site histidine residue in Escherichia coli thioesterase I/protease I/lysophospholipase L 1

Wen Jin Wu, Sergiy I. Tyukhtenko, Tai-huang Huang

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Abstract

Owing to the hydrogen-bond interaction and rapid exchange rate with the bulk water, the transverse relaxation time for the N δ1 -H proton of the catalytic histidine in Escherichia coli thioesterase I/protease I/lysophospholipase L 1 (TEP-I) is rather short. Because of its catalytic importance, it is desirable to detect and assign this proton resonance. In this paper, we report the first direct NMR correlation between the short-lived N δ1 -H proton and its covalently attached N δ1 -nitrogen of the catalytic His157 residue in E. coli thioesterase/protease I. We have used gradient-enhanced jump-return spin-echo HMQC (GE-JR SE HMQC) to obtain a direct correlation between the short-lived N δ1 -H proton and its covalently attached N δ1 - nitrogen. The sensitivity of detection for the short-lived N δ1 - H proton was enhanced substantially by improved water suppression, in particular, the suppression of radiation damping via pulsed field gradients.

Original languageEnglish
Pages (from-to)1037-1040
Number of pages4
JournalMagnetic Resonance in Chemistry
Volume44
Issue number11
DOIs
Publication statusPublished - 2006 Nov 1

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Keywords

  • Catalytic triad
  • Jump-and-return HMQC
  • Low-field resonance
  • NMR
  • Protease
  • Strong hydrogen bond
  • Thioesterase

ASJC Scopus subject areas

  • Chemistry(all)
  • Materials Science(all)

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