PMID: 9421528Feb 28, 1998Paper

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

Nucleic Acids Research
D L WongN O Reich

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.

References

Jul 1, 1978·Nucleic Acids Research·P J GreeneH W Boyer
Mar 6, 1992·Biochemistry·N O Reich, M J Danzitz
Mar 19, 1991·Biochemistry·N O Reich, N Mashhoon
Jul 15, 1981·Journal of Molecular Biology·R E Dickerson, H R Drew
May 24, 1984·Nature·C A FrederickJ M Rosenberg
Jun 1, 1983·Photochemistry and Photobiology·R O Rahn, H G Sellin
Nov 8, 1994·Proceedings of the National Academy of Sciences of the United States of America·J LabahnW Saenger
Jul 23, 1996·Proceedings of the National Academy of Sciences of the United States of America·R A GarciaN O Reich
Oct 1, 1996·Nucleic Acids Research·O N JensenD F Barofsky
Nov 26, 1996·Biochemistry·B W Allan, N O Reich

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Citations

Dec 12, 2002·Mass Spectrometry Reviews·Hanno Steen, Ole Nørregaard Jensen
Jun 1, 2000·Journal of the American Society for Mass Spectrometry·R A RiegerC R Iden
Jul 30, 2010·Journal of Proteome Research·Erin D Michaelson-RichieNatalia Y Tretyakova
Jan 13, 2000·Protein Science : a Publication of the Protein Society·M C GoldenT H Koch
Mar 11, 2003·Microbiology and Molecular Biology Reviews : MMBR·Eric S MillerWolfgang Rüger
Jul 20, 2005·BMC Bioinformatics·Yong TangSteven F Jennings
Oct 4, 2014·PloS One·Helen E BarnesR Allyn Forsyth
Apr 3, 1999·The Journal of Biological Chemistry·V PingoudA Pingoud
Feb 8, 2002·The Journal of Biological Chemistry·Merlind MückeMonika Reuter
Jun 26, 1999·The Journal of Biological Chemistry·B W AllanN O Reich

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