Nucleic Acids Research, Vol 24, Issue 19 3866-3872, Copyright © 1996 by Oxford University Press
ON Jensen, S Kulkarni, JV Aldrich and DF Barofsky
Two peptide-oligothymidylic acids, prepared by joining an 11 residue
synthetic peptide containing one internal carboxyl group (Asp side chain)
to amino-linker-5'pdT6 and amino-linker-5'pdT10 oligonucleotides, were
analyzed by matrix-assisted laser desorption/ionization (MALDI) on a linear
time-of-flight mass spectrometer and by electrospray ionization (ESI) on a
triple- quadrupole system. These synthetic compounds model peptide-nucleic
acid heteroconjugates encountered in antisense research and in studies that
use photochemical crosslinking to investigate molecular aspects of
protein-nucleic acid interactions. MALDI and ESI sensitivities for the two
hybrid compounds were found to be similar respectively to their
sensitivities for the pure oligonucleotide parts. In general, MALDI proved
to be less affected by sample impurities and more sensitive than ESI, while
ESI on the quadrupole produced greater mass accuracy and resolution than
MALDI on the time-of-flight instrument. A hybrid's behavior in a
MALDI-matrix or an ESI-spray-solvent was found to be governed mainly by the
oligonucleotide. A single positive ESI tandem mass spectrum of the
peptide-dT6 accounted for the heteroconjugate's entire primary structure
including the point of the oligonucleotide's covalent attachment to the
peptide.
ARTICLES
Characterization of peptide-oligonucleotide heteroconjugates by mass spectrometry
Department of Biochemistry and Biophysics, College of Pharmacy, Oregon State University, Corvallis 97331-7301, USA.
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