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Glutarate And N-Acetyl-L-Glutamate Buffers For Cell-Free Synthesis Of Selectively 15n-Labelled Proteins, X Jia, Kiyoshi Ozawa, K Loscha, G Otting Jan 2009

Glutarate And N-Acetyl-L-Glutamate Buffers For Cell-Free Synthesis Of Selectively 15n-Labelled Proteins, X Jia, Kiyoshi Ozawa, K Loscha, G Otting

Faculty of Science - Papers (Archive)

Cell-free protein synthesis provides rapid and economical access to selectively 15N-labelled proteins, greatly facilitating the assignment of 15N-HSQC spectra. While the best yields are usually obtained with buffers containing high concentrations of potassium L-glutamate, preparation of selectively 15N-Glu labelled samples requires non-standard conditions. Among many compounds tested to replace the L-Glu buffer, potassium N-acetyl-L-glutamate and potassium glutarate were found to perform best, delivering high yields for all proteins tested, with preserved selectivity of 15N-Glu labelling. Assessment of amino-transferase activity by combinatorial 15N-labelling revealed that glutarate and N-acetyl-L-glutamate suppress the transfer of the 15N-alpha-amino groups between amino acids less well than …


Positionally Dependent 15n Fractionation Factors In The Uv Photolysis Of N2o Determined By High Resolution Ftir Spectroscopy, Fred Turatti, D W. T Griffith, Stephen Wilson, Michael Esler, T Rahn, H Zang, G A. Blake Jan 2000

Positionally Dependent 15n Fractionation Factors In The Uv Photolysis Of N2o Determined By High Resolution Ftir Spectroscopy, Fred Turatti, D W. T Griffith, Stephen Wilson, Michael Esler, T Rahn, H Zang, G A. Blake

Faculty of Science, Medicine and Health - Papers: part A

Positionally dependent fractionation factors for the photolysis of isotopomers of N2O in natural abundance have been determined by high resolution FTIR spectroscopy at three photolysis wavelengths. Fractionation factors show clear 15N position and photolysis wavelength dependence and are in qualitative agreement with theoretical models but are twice as large. The fractionation factors increase with photolysis wavelength from 193 to 211 nm, with the fractionation factors at 207.6 nm for 14N15N16O, 15N14N16O and 14N14N18O equal to −66.5±5‰,−27.1±6‰ and −49±10‰, respectively.