Identification of a new gene product of PKIbeta by HPLC-ESIMS.
An identification method using high-performance liquid chromatography combined with electrospray mass spectrometry (HPLC-ESIMS) has been developed to verify an expressed gene product of kinase inhibitor (PKIbeta). This protein was expressed in this university for the first time from a newly cloned gene in the cDNA library of human fetal brain. The measured MW (8468.9 Da) of PKIbeta-78 was consistent with expectations. The gene product of PKIbeta-78 was monitored by ESIMS to ensure there was no mis-expressed PKIbeta-70 in the process of gene engineering. The peptide mapping of PKIbeta-78 and its partial sequence were, furthermore, determined. By database searching based on the experimental MWs and partial sequences provided, it was verified that this gene product is a new protein. The pseudosubstrate site and leucine-rich site for the function region of PKIbeta-78 are also confirmed.
Song
H
H
State Key Laboratory of Genetic Engineering, Fudan University, Shanghai, China.
Yue
G
G
Zhou
Z
Z
Xie
Y
Y
Yang
P
P
Mao
Y
Y
Xu
Y
Y
Wang
H
H
D000595:Amino Acid Sequence; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D018076:DNA, Complementary; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010449:Peptide Mapping; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D014357:Trypsin / Q000737:chemistry
D002352:Carrier Proteins; D018076:DNA, Complementary; D047908:Intracellular Signaling Peptides and Proteins; C015482:protein kinase modulator; D014357:Trypsin