USE OF HYDROPHILIC INTERACTION CHROMATOGRAPHY FOR THE STUDY OF TYROSINE PROTEIN-KINASE SPECIFICITY

被引:30
作者
BOUTIN, JA [1 ]
ERNOULD, AP [1 ]
FERRY, G [1 ]
GENTON, A [1 ]
ALPERT, AJ [1 ]
机构
[1] POLYLC INC,COLUMBIA,MD 21045
来源
JOURNAL OF CHROMATOGRAPHY-BIOMEDICAL APPLICATIONS | 1992年 / 583卷 / 02期
关键词
D O I
10.1016/0378-4347(92)80546-3
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
A new HPLC method has been developed to assay tyrosine protein kinase activity. Using hydrophilic interaction chromatography, it is possible to resolve the four components of the incubation medium: substrate peptide, [P-32]phosphorylated peptide, unreacted [gamma-P-32]ATP, and P-32-labelled inorganic phosphate. ATP interacts so strongly with the stationary phase material that it can be removed selectively from the incubation medium with solid-phase extraction cartridges packed with the same type of material. The three remaining components of interest can then be resolved by reversed-phase or hydrophilic interaction HPLC. This procedure permits the evaluation of almost every type of peptide as a substrate of tyrosine protein kinase.
引用
收藏
页码:137 / 143
页数:7
相关论文
共 19 条
[1]   SYNTHETIC PEPTIDES AS MODEL SUBSTRATES FOR THE STUDY OF THE SPECIFICITY OF THE POLYCATION-STIMULATED PROTEIN PHOSPHATASES [J].
AGOSTINIS, P ;
GORIS, J ;
PINNA, LA ;
MARCHIORI, F ;
PERICH, JW ;
MEYER, HE ;
MERLEVEDE, W .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1990, 189 (02) :235-241
[2]   HYDROPHILIC-INTERACTION CHROMATOGRAPHY FOR THE SEPARATION OF PEPTIDES, NUCLEIC-ACIDS AND OTHER POLAR COMPOUNDS [J].
ALPERT, AJ .
JOURNAL OF CHROMATOGRAPHY, 1990, 499 :177-196
[3]  
ALPERT AJ, UNPUB
[4]  
BARRET JM, 1992, IN PRESS CHEM BIOL I
[5]   PARTIAL-PURIFICATION AND CHARACTERIZATION OF A NEW P36/40 TYROSINE PROTEIN-KINASE FROM HL-60 [J].
BOUTIN, JA ;
ERNOULD, AP ;
GENTON, A ;
CUDENNEC, CA .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1989, 160 (03) :1203-1211
[6]   A RAPID ASSAY FOR PROTEIN-KINASES PHOSPHORYLATING SMALL POLYPEPTIDES AND OTHER SUBSTRATES [J].
BRAUN, S ;
GHANY, MA ;
RACKER, E .
ANALYTICAL BIOCHEMISTRY, 1983, 135 (02) :369-378
[7]   PHOSPHORYLATION OF SMALL PEPTIDES BY SPLEEN TPK-IIA, A TYROSINE PROTEIN-KINASE STIMULATED BY POLYLYSINE AND BY HIGH IONIC-STRENGTH [J].
BRUNATI, AM ;
MARCHIORI, F ;
RUZZA, P ;
CALDERAN, A ;
BORIN, G ;
PINNA, LA .
FEBS LETTERS, 1989, 254 (1-2) :145-149
[8]   CHARACTERIZATION OF 4 TYROSINE PROTEIN-KINASES FROM THE PARTICULATE-FRACTION OF RAT SPLEEN [J].
BRUNATI, AM ;
PINNA, LA .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1988, 172 (02) :451-457
[9]   SYNTHETIC PEPTIDES REPRODUCING THE EGF-RECEPTOR SEGMENT HOMOLOGOUS TO THE PP60V-SRC PHOSPHOACCEPTOR SITE - PHOSPHORYLATION BY TYROSINE PROTEIN-KINASES [J].
COLA, C ;
BRUNATI, AM ;
BORIN, G ;
RUZZA, P ;
CALDERAN, A ;
DECASTIGLIONE, R ;
PINNA, LA .
BIOCHIMICA ET BIOPHYSICA ACTA, 1989, 1012 (02) :191-195
[10]  
ERICKSON RL, 1979, P NATL ACAD SCI USA, V76, P6260