Biological fingerprinting analysis of the interactome of a kinase inhibitor in human plasma by a chemiproteomic approach.
In this study, a gel free chemiproteomic method based on chromatography was developed and applied for the biological fingerprinting analysis of complex biological system. p-Aminobenzamidine (ABA), an inhibitor of trypsin-like serine proteases, was immobilized for characterizing their interacting proteins in human plasma. By the proteomic analysis method, 214 proteins were identified with obvious affinity to the immobilized ABA. By searching the sequences of above proteins with consensus patterns of the two active sites, seven proteins belong to trypsin-like serine protease group were found. Based on the Gene Ontology annotation, the identified trypsin-like serine proteases have the function of catalytic activity and calcium ion binding, and are mainly involved in the biological process of blood coagulation. Eight more other proteins related to calcium ion binding and blood coagulation were found. Nearly all of these proteins cannot be identified by directly analyzing the plasma sample demonstrating the chemiproteomics a useful approach to characterize interacting proteins in the low abundance range.
Tian
Ruijun
R
National Chromatographic R&A Center, Dalian Institute of Chemical Physics, The Chinese Academy of Sciences, Dalian 116023, China.
Jiang
Xinning
X
Li
Xin
X
Jiang
Xiaogang
X
Feng
Shun
S
Xu
Songyun
S
Han
Guanghui
G
Ye
Mingliang
M
Zou
Hanfa
H
D000328:Adult; D001550:Benzamidines / Q000737:chemistry; D001777:Blood Coagulation; D001798:Blood Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000737:chemistry; D002846:Chromatography, Affinity; D006801:Humans; D008297:Male; D010449:Peptide Mapping / Q000379:methods*; D047428:Protein Kinase Inhibitors / Q000097:blood* / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D012710:Serum Albumin, Bovine / Q000302:isolation & purification; D053719:Tandem Mass Spectrometry; D014357:Trypsin / Q000302:isolation & purification
D001550:Benzamidines; D001798:Blood Proteins; D002135:Calcium-Binding Proteins; D047428:Protein Kinase Inhibitors; D012710:Serum Albumin, Bovine; C006500:4-aminobenzamidine; D014357:Trypsin