The stereochemical course of the wheat germ meso-diaminopimelate (DAP) decarboxylase reaction is compared to that of the decarboxylase isolated from Bacillus sphaericus, which has been reported to proceed with an unusual inversion of configuration [Asada, Y., Tanizawa, K., Sawada, S., Suzuki, T., Misono, H., & Soda, K. (1981) Biochemistry 20, 6881-6886]. Reaction of each enzyme with either unlabeled diaminopimelic acid in D2O or [2,6-2H2]diaminopimelic acid in H2O gave stereospecifically deuterium-labeled lysine samples that were derivatized with (-)-camphanoyl chloride and diazomethane. Analysis by two-dimensional 1H-13C heteronuclear NMR shift correlation spectroscopy with 2H decoupling confirmed the stereochemistry of the B. sphaericus enzyme reaction and showed that the eukaryotic wheat germ meso-DAP decarboxylase also operates with inversion of configuration. This suggests similar mechanisms for the prokaryotic and eukaryotic enzymes and contrasts the retention mode observed with other pyridoxal phosphate dependent alpha-decarboxylases.
The fast-atom-bombardment mass spectra (FAB-MS) of a series of ribo-and deoxyribonucleosides and -nucleotides were examined in both positive-ion and negative-ion modes. Behaviour of cytosine analogs having unnatural bases (2( 1H)-pyrimidone, 2( 1H)-pyridinone, and 4-amino-2( 1 H)-pyridinone) was similar to that of compounds having naturally occurring bases. Nucleotides protected by dimethoxytrityl and p-chlorophenyl groups were also investigated using this technique. Use of negative-ion mode reduces interference from positive counter-ions (e.g., Na+) and allows rapid sequence detennindtion of simple di-and trinucleotides.Introduction. -Mass spectrometry possesses great potential as a technique for the study of biological molecules, because only small amounts of sample are required of materials which may be available only in minute quantities [2]. However, in the past, its use has been hindered by the nonvolatility of large polar molecules and by their sensitivity to the extreme conditions needed to bring them into the gas phase [3]. The advent of milder desorption ionization methods, including fast-atom bombardment has solved this problem for many types of naturally occurring molecules [4].Fast-atom-bombardment mass spectrometry (FAB-MS) [5] is a technique, whereby a stream of fast atoms (typically Ar or Xe) impinges onto a solution of the sample to be analysed in a liquid matrix such as glycerol. Volatilization and ionization [6] of the analyte occurs, and the resulting ions, either positive or negative can be detected in the usual way. The 'sputtering' or desorption of the analyte into the gas phase is most efficient, when the fast atom beam strikes the surface of the glycerol at approximately a 70" angle of incidence [7].The study of nucleosides and nucleotides by MS has been difficult because of the involatility and thermal instability of these compounds. However, recently, other 'soft' ionization methods [8-101 such as field desorption [ 1 11, californium-252-plasma desorption [ 121, secondary-ion MS [ 131, pulsed-laser-induced desorption [ 141, and atmosphericpressure-ionization MS [ 151 as well as pyrolysis-electron-impact and chemical-ionization MS [16] have been applied successfully in this area. These methods have also been used to
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