Click chemistry generated model DNA-peptide heteroconjugates as tools for mass spectrometry

Analytical Chemistry
Fiona J FlettHeidrun Interthal

Abstract

UV cross-linking of nucleic acids to proteins in combination with mass spectrometry is a powerful technique to identify proteins, peptides, and the amino acids involved in intermolecular interactions within nucleic acid-protein complexes. However, the mass spectrometric identification of cross-linked nucleic acid-protein heteroconjugates in complex mixtures and MS/MS characterization of the specific sites of cross-linking is extremely challenging. As a tool for the optimization of sample preparation, ionization, fragmentation, and detection by mass spectrometry, novel synthetic DNA-peptide heteroconjugates were generated to act as mimics of UV cross-linked heteroconjugates. Click chemistry was employed to cross-link peptides to DNA oligonucleotides. These heteroconjugates were fully characterized by high resolution FTICR mass spectrometry and by collision-induced dissociation (CID) following nuclease P1 digestion of the DNA moiety to a single nucleotide monophosphate. This allowed the exact site of the cross-linking within the peptide to be unambiguously assigned. These synthetic DNA-peptide heteroconjugates have the potential to be of use for a variety of applications that involve DNA-peptide heteroconjugates.

References

Oct 1, 1996·Nucleic Acids Research·O N JensenD F Barofsky
Dec 12, 2002·Mass Spectrometry Reviews·Hanno Steen, Ole Nørregaard Jensen
Mar 6, 2008·Journal of Mass Spectrometry : JMS·Soheil Pourshahian, Patrick A Limbach
May 4, 2010·Journal of the American Society for Mass Spectrometry·Kady L Krivos, Patrick A Limbach
Feb 9, 2011·The Journal of Organic Chemistry·Takeshi YamadaKallanthottathil G Rajeev
Mar 28, 2012·Journal of the American Society for Mass Spectrometry·Brittany CatronPatrick A Limbach
May 12, 2012·Journal of Proteomics·Carla SchmidtHenning Urlaub
May 18, 2013·Organic & Biomolecular Chemistry·I Kira AstakhovaJesper Wengel
Aug 1, 1994·Journal of the American Society for Mass Spectrometry·S A McLuckey, S Habibi-Goudarzi
Feb 1, 1995·Journal of the American Society for Mass Spectrometry·S Habibi-Goudarzi, S A McLuckey
Jan 1, 1992·Journal of the American Society for Mass Spectrometry·S A McLuckeyG L Glish
Nov 21, 2013·Current Medicinal Chemistry·Chang-Fang WangHélder A Santos
Jun 12, 2014·ACS Chemical Biology·Jung Eun YeoNatalia Y Tretyakova

❮ Previous
Next ❯

Citations

Jan 4, 2018·Nature Communications·Fiona J FlettJulia M Richardson
Apr 5, 2019·Journal of the American Society for Mass Spectrometry·Yang Liu, František Tureček
Oct 11, 2017·Macromolecular Rapid Communications·Steven MartensFilip E Du Prez
Jan 17, 2017·Journal of the American Society for Mass Spectrometry·Pengfei Li, Glen P Jackson

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.