Author: Zhou, ShengTao; Liu, Rui; Zhao, Xia; Huang, CanHua; Wei, YuQuan
Title: Viral proteomics: The emerging cutting-edge of virus research Document date: 2011_6_26
ID: 3ahamzjv_44
Snippet: Among the various stable isotopic labeling methods, the newly established 18 O-labeling is the most cost-effective, in that the labeling reagent is heavy water (D 2 O). However, the major limitation of this approach lies in the fact that the analysis of the low mass difference between peptides, typically 2-4 atomic mass units, requires a highly sensitive MS instrument [58] . In a recent report, 18 O-labeling was used to study host-virus interacti.....
Document: Among the various stable isotopic labeling methods, the newly established 18 O-labeling is the most cost-effective, in that the labeling reagent is heavy water (D 2 O). However, the major limitation of this approach lies in the fact that the analysis of the low mass difference between peptides, typically 2-4 atomic mass units, requires a highly sensitive MS instrument [58] . In a recent report, 18 O-labeling was used to study host-virus interactions between HIV-1 and the HIV-infected CD4 + cell line CEMx174. The cellular proteome of CEMx174 cell line upon HIV-1 infection was assessed using stable isotope labeling, 18 O/ 16 O, and accurate mass and time (AMT) tagging. The 18 O/ 16 O ratio was subsequently quantified with a highly sensitive FTICR-MS instrument. 3255 unique host proteins were identified, of which 344 were upregulated and 343 were downregulated. Functional analysis unraveled changes in proteins from various functional categories, including nuclear transport, ubiquitination, cell cycle progression, and the citrate cycle. At least seven proteins (CASP3, DLD, KPNA3, KPNB1, RAN, TPR, and UBE1) were further validated by Western blotting [59] .
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