Nucleic Acids Research 1998-01-15

Electrospray ionization mass spectrometric characterization of photocrosslinked DNA-EcoRI DNA methyltransferase complexes.

D L Wong, J G Pavlovich, N O Reich

Index: Nucleic Acids Res. 26(2) , 645-9, (1998)

Full Text: HTML

Abstract

We describe a novel strategy combining photocrosslinking and HPLC-based electrospray ionization mass spectrometry to identify UV crosslinked DNA-protein complexes. Eco RI DNA methyltransferase modifies the second adenine within the recognition sequence GAATTC. Substitution of 5-iodouracil for the thymine adjacent to the target base (GAATTC) does not detectably alter the DNA-protein complex. Irradiation of the 5-iodouracil-substituted DNA-protein complex at various wavelengths was optimized, with a crosslinking yield >60% at 313 nm after 1 min. No protein degradation was observed under these conditions. The crosslinked DNA-protein complex was further analyzed by electrospray ionization mass spectrometry. The total mass is consistent with irradiation-dependent covalent bond formation between one strand of DNA and the protein. These preliminary results support the possibility of identifying picomole quantities of crosslinked peptides by similar strategies.

Related Compounds

  • 5-Iodouracil

Related Articles:

Enzymatic primer-extension with glycerol-nucleoside triphosphates on DNA templates.

2009-01-01

[PLoS ONE 4(3) , e4949, (2009)]

Highly hydrophilic and nonionic poly(2-vinyloxazoline)-grafted silica: a novel organic phase for high-selectivity hydrophilic interaction chromatography.

2014-07-01

[Anal. Bioanal. Chem 406(19) , 4585-93, (2014)]

Preparation and properties of oligodeoxynucleotides containing 5-iodouracil and 5-bromo- and 5-iodocytosine.

1997-01-01

[Bioconjug. Chem. 8(5) , 757-61, (1997)]

A simple crosslinking method, CLAMP, to map the sites of RNA-contacting domains within a protein.

2008-01-01

[Methods Mol. Biol. 488 , 181-90, (2008)]

Photoreactivity of 5-iodouracil-containing DNA-Sso7d complex.

1999-01-01

[Nucleic Acids Symp. Ser. (42) , 171-2, (1999)]

More Articles...