Development and
Preliminary Application of a Peptide Mass Fingerprinting Technique in Proteome
Research
WANG Jing-Lan1, WAN Jing-Hong2, LUO
Ling2, XIONG Shao-Xiang3, WANG Guang-Hui3, HE
Fu-Chu2, QIAN Xiao-Hong1*
( 1National Center of Biomedical Analysis, Beijing 100850； 2Institute of Radiation Medicine, Chinese
Academy of Military Science, Beijing 100850； 3National Center of Mass Spectrometry,
Beijing 100080,
China )
Abstract
Proteome means the total proteins expressed by the genome in a cell or tissue.
Two-dimensional electrophoresis (2-DE) is now the most powerful separating
technique and the key separation method used in proteome. Peptide mass
fingerprinting (PMF) is becoming a widely used and vastly developed technique
for protein identification in 2-D gels.
In this research, a systematic method to identify the proteins in
polyacrylamide gels by PMF was developed. Proteins were digested in-gel by
enzyme and the masses of generated peptides were measured by matrix assisted
laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS).
The data obtained from PMF were used in protein database search and the protein
identification.
The proteins from human lung cancer cells isolated by 2-DE were subjected to
identification by the PMF method developed in this work. Three spots of
proteins in gel were identified as G3P2_HUMAN, UBL1_ HUMAN and TPIS_HUMAN.
Key words proteome； peptide mass fingerprinting
(PMF)；
two dimensional electrophoresis； human lung cancer cell； matrix assisted laser
desorption/ionization time of flight mass spectrometry (MALDI-TOF MS)
*Corresponding author： Tel, 86-10-66930306； Fax, 86-10-68186281； e-mail, [email protected]