Characterization of peptide-oligonucleotide heteroconjugates by mass spectrometry.
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.
Jensen
O N
ON
Department of Biochemistry and Biophysics, College of Pharmacy, Oregon State University, Corvallis 97331-7301, USA.
Kulkarni
S
S
Aldrich
J V
JV
Barofsky
D F
DF
D000595:Amino Acid Sequence; D013058:Mass Spectrometry / Q000379:methods; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*
D009841:Oligonucleotides; D010455:Peptides