1. Basic Information

Name:RASH_HUMAN
Accession#:P01112
Description:GTPase HRas
AA Number:189
Sequence:
1
51
101
151
   MTEYKLVVVG AGGVGKSALT IQLIQNHFVD EYDPTIEDSY RKQVVIDGET
CLLDILDTAG QEEYSAMRDQ YMRTGEGFLC VFAINNTKSF EDIHQYREQI
KRVKDSDDVP MVLVGNKCDL AARTVESRQA QDLARSYGIP YIETSAKTRQ
GVEDAFYTLV R
EIRQHKLRK LNPPDESGPG CMSCKCVLS
*Highlighted peptides (with yellow background) have developed assays.
*Green background amino acids are PTMs.

2. Protein Separation

Sample Preparation: Cells from Melanoma cancer lines were lysed in 8M urea/ 100mM ammonium bicarbonate buffer on ice. The equivalent of 200,000 cells were loaded onto a SDS gel. The protein of interest was excised and ingel digestion with trypsin was performed after reduction and alkylation with TCEP and IAA. 1/6 of the resulting digest was then analyzed on a Thermo Scientific TSQ mass spectrometer.

 

3. LC-MS/MS Data

LVVVGAGGVGK

    No LC-MS/MS for this peptide.

SYGIPYIETSAK

    No LC-MS/MS for this peptide.

QGVEDAFYTLVR

    No LC-MS/MS for this peptide.

4. LC-MRM Screening

Peptides screening: Peptides and transitions for analysis were selected using a predictive peptide program. The results were screened for those with the highest specificity and the most intense transitions.