Development of a novel quadrupole ion trap time-of-flight mass spectrometer and cryogenic ion spectroscopy for identification of phosphotyrosine

Author(s)
권장한
Advisor
강혁
Department
일반대학원 에너지시스템학과
Publisher
The Graduate School, Ajou University
Publication Year
2023-08
Language
eng
Keyword
cryogenic spectroscopymass spectrometertryptic peptide
Alternative Abstract
We constructed a novel QIT-TOF mass spectrometer. We obtained room temperature UV PD spectra at the home of singly protonated tyrosine and singly protonated hexapeptides, DYYVVR. DYYVVR is a tryptic peptide of Janus kinase 3 and has the two active tyrosine residues needed to regulate of the protein’s activity by phosphorylation. Although we obtained a well-resolved spectrum at room temperature for singly protonated tyrosine, two adjacent tyrosine absorption spectra in singly protonated DYYVVR were not distinguishable. Therefore, cryogenic ion spectroscopy (CIS) was used to elucidate the spectrum results of singly protonated DYYVVR at room temperature. Through comparison with point mutants, it was determined that the lower electronic band to the absorption of 981st tyrosine, while the higher band to 980th tyrosine. In the case of phosphorylation, the UV absorption of the phosphorylated chromophore shifts to higher energy above 36500 cm–1. In contrast, the unphosphorylated chromophore retains its absorption in the same region. It is proved that CIS can be used to identify the two adjacent tyrosine chromophores and to confirm the phosphorylation site of a kinase homology domain. In the case of room temperature, UVPD spectra are also used to differentiate the absorption of two adjacent tyrosine chromophores. Mass spectrometry and ultraviolet spectroscopy in 2-dimensional analysis can effectively demonstrate to identify the location of phosphorylation sites in a peptide.
URI
https://dspace.ajou.ac.kr/handle/2018.oak/24337
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Graduate School of Ajou University > Department of Energy Systems > 4. Theses(Ph.D)
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