Participants/Project Abstracts
Summer 2007 Bioanalytical Science REU Participants
EXPLORING
THE FUNDAMENTALS OF SNAPP MASS SPECTROMETRY
Christine E. Reder, Jolene K. Diedrich, Ryan R.
Julian
Department
of Chemistry, University of California – Riverside, UCR 92521 and Ferris
State University, Big Rapids, MI 49307.
Abstract
The focus of this project is to continue the
development of the SNAPP technique, which can be used to observe protein
structure using mass spectrometry. Previous work with 18-crown-6 ether (18C6) has shown protein structure
can be examined through the availability of amino acid side-chains, determined
through the interaction of lysine and 18C6. Here, the availability of arginine side-chains is explored
though interactions with other crown ethers, as analyzed through the use of
electrospray ionization mass spectrometry (ESI-MS). Both, dibenzo-30-crown-10 ether (DB30C10) and DP (a crown
ether which contains two endocyclic dialkylhydrogenphosphate esters)
preferentially form noncovalent complexes with the guanidinium group of the
arginine side-chain. Experiments
were conducted using various solvent conditions for each crown with
ubiquitin. Also, varying
electrospray parameters were used for the ESI-MS analysis of ubiquitin and 18C6
solutions to observe the effects of the different settings on crown attachment. DB30C10 shows a significant solvent
effect on crown attachment with the addition of organic co-solvent to the
solution. DP binds to protein with
an uneven distribution of crown among the charge states. The 18C6 solutions illustrate the
importance of keeping instrument settings constant through the course of an
experiment because changes in the settings affect the number of crowns
attaching.
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