A rapid isolation and identification method for blocked N-terminal peptides by isothiocyanate-coupled magnetic nanoparticles and MS.
A quick isolation and identification of N-blocked peptides from protein digest mixtures were achieved by diisothiocyanate or isothiocyanate-coupled magnetic nanoparticles and MS. After protein digests were guanidinated and then mixed with diisothiocyanate or isothiocyanate-coupled magnetic nanoparticles, unmodified N-terminal peptides were covalently bound to magnetic nanoparticles, and can be removed from the mixture under magnetic field. Therefore, N-blocked peptides could be isolated and analyzed by MALDI or ESI MS. This new strategy was demonstrated with model peptides, proteins, and the lysates of HepG2 cells.
Zhao
Liyan
L
State Key Lab of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing, P. R. China.
Zhang
Yangjun
Y
Wei
Junying
J
Cao
Dong
D
Liu
Kehui
K
Qian
Xiaohong
X
D045744:Cell Line, Tumor; D002852:Chromatography, Ion Exchange; D052203:Ferrosoferric Oxide / Q000737:chemistry*; D006801:Humans; D017879:Isothiocyanates / Q000737:chemistry*; D053768:Metal Nanoparticles / Q000737:chemistry*; D046529:Microscopy, Electron, Transmission; D010455:Peptides / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013997:Time Factors
D017879:Isothiocyanates; D010455:Peptides; C037152:isothiocyanic acid; D052203:Ferrosoferric Oxide