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| Type |
Poster Presentation |
| Area |
Analytical Chemistry |
| Room No. |
Event Hall |
| Time |
4월 19일 (목요일) 11:00~12:30 |
| Code |
ANAL.P-291 |
| Subject |
Comprehensive analysis of the RNAlater effect on global proteome and phosphoproteome of human pancreatic tumor tissues |
| Authors |
jingi Bae, Su-Jin Kim, Sang-Won Lee*
Department of Chemistry, Korea University, Korea
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| Abstract |
Pancreatic tumor tissues contain a variety of enzymes including DNases, RNases and proteases which may lead to autolysis during sample processing. In general, RNAlater can be used to keep the molecules (mainly, nucleic acids) intact since it contains high concentrations of quaternary ammonium sulfates which denature, for example, RNases and DNases. However, it is largely known whether RNAlater affect the qualitative and quantitative information of proteome and phosphoproteome. Thus, we have carried out a systematic and comprehensive analysis of the RNAlater effect on the proteome and phosphoproteome..
Pancreatic tumor tissues from three patients were pulverized using Covaris CP02 Cryoprep device and then half of powders was stored at -80 °C (FF) and the other half was incubated in RNAlater at 4 °C for 24 hours. We used the 6-plex TMT for labeling on the peptide after modified FASP (Filter Aided Sample Preparation) for digestion. Labeled sample were separated into 24 fractions by mid pH reverse phase liquid chromatography. Then, a portion (~8%) of each mRP fraction was subjected to LC-MS/MS to profile the global proteome and the rest (~92%) was concatenated into 12 fractions, each of which were then performed IMAC–based phosphopeptide enrichment. As a result, the global profiling identified 186,941 unmodified peptides of 9,152 protein groups, 18,705 phosphopeptides (15,842 phosphosites). We have observed the narrow distribution of quantitative changes in phosphorylation between the FF and RNAlater. Therefore, it was confirmed that stored tissues in RNAlater do not significantly affect the proteome and phosphoproteome of pancreatic cancer tumors.
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| E-mail |
bjg0926@nate.com |
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121st General Meeting of the KCS