DETECTION OF NONCOVALENT FKBP FK506 AND FKBP RAPAMYCIN COMPLEXES BY CAPILLARY ELECTROPHORESIS MASS-SPECTROMETRY AND CAPILLARY ELECTROPHORESIS TANDEM MASS-SPECTROMETRY

被引：31

作者：

HSIEH, YL ^{[1
]}

CAI, JY ^{[1
]}

LI, YT ^{[1
]}

HENION, JD ^{[1
]}

GANEM, B ^{[1
]}

机构：

[1] CORNELL UNIV, BAKER LAB, DEPT CHEM, 927 WARREN DR, ITHACA, NY 14850 USA

来源：

JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY | 1995年 / 6卷 / 02期

基金：

美国国家卫生研究院;

关键词：

D O I：

10.1016/S1044-0305(94)00097-J

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The well known biospecific noncovalent receptor-ligand association complexes between the immunophilin FKBP and the immunosuppressive drugs FK506 and Rapamycin (RM) were investigated by on-line capillary electrophoresis-mass spectrometry (CE-MS) under selected ion monitoring (SIM) conditions and by CE-MS with tandem mass spectrometry (CE-MS/MS) under selected reaction monitoring (SRM) conditions. Solutions of hFKBP (33.3 muM) were dissolved in 50 mM ammonium acetate at pH 7.5. Samples that contained 100 muM of FK506 or RM also were prepared under the same solution conditions. By using these aqueous pH neutral conditions, samples were analyzed by SIM CE-MS and SRM CE-MS and the target complexes were separated by CE with mass spectrometer detection of the individual complexes between FKBP and FK506 [hFKBP + FK506 + 7H]7+ as well as FKBP and RM [hFKBP + RM + 7H]7+. In an experiment where a mixture of FK506 and RM was analyzed in the presence of FKBP, a nine-to-one ratio of ion current abundances between the RM and FK506 complexes was observed as reported in the literature from other studies. These results suggest that CE-MS and CE-MS/MS may be yet another analytical method for studying noncovalent interactions of biologically important macromolecules under physiological conditions.

引用

页码：85 / 90

页数：6

共 40 条

[1] DIRECT OBSERVATION OF A TERNARY COMPLEX BETWEEN THE DIMERIC ENZYME HIV-1 PROTEASE AND A SUBSTRATE-BASED INHIBITOR
BACA, M
KENT, SBH
[J]. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1992, 114 (10) : 3992 - 3993
[2] CONTRIBUTIONS OF MASS-SPECTROMETRY TO PEPTIDE AND PROTEIN-STRUCTURE
BIEMANN, K
[J]. BIOMEDICAL AND ENVIRONMENTAL MASS SPECTROMETRY, 1988, 16 (1-12): : 99 - 111
[3] 2 DISTINCT SIGNAL TRANSMISSION PATHWAYS IN LYMPHOCYTES-T ARE INHIBITED BY COMPLEXES FORMED BETWEEN AN IMMUNOPHILIN AND EITHER FK506 OR RAPAMYCIN
BIERER, BE
MATTILA, PS
STANDAERT, RF
HERZENBERG, LA
BURAKOFF, SJ
CRABTREE, G
SCHREIBER, SL
[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (23) : 9231 - 9235
[4] OBSERVATION OF NONCOVALENT ENZYME SUBSTRATE AND ENZYME PRODUCT COMPLEXES BY ION-SPRAY MASS-SPECTROMETRY
GANEM, B
LI, YT
HENION, JD
[J]. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1991, 113 (20) : 7818 - 7819
[5] DETECTION OF OLIGONUCLEOTIDE DUPLEX FORMS BY ION-SPRAY MASS-SPECTROMETRY
GANEM, B
LI, YT
HENION, JD
[J]. TETRAHEDRON LETTERS, 1993, 34 (09) : 1445 - 1448
[6] DETECTION OF NONCOVALENT RECEPTOR LIGAND COMPLEXES BY MASS-SPECTROMETRY
GANEM, B
LI, YT
HENION, JD
[J]. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1991, 113 (16) : 6294 - 6296
[7] ANALYSIS OF THE STOICHIOMETRY OF THE T4 GENE 45 PROTEIN BY ION-SPRAY MASS-SPECTROMETRY
GANEM, B
LI, YT
HSIEH, YL
HENION, JD
KABOORD, BF
FREY, MW
BENKOVIC, SJ
[J]. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1994, 116 (04) : 1352 - 1358
[8] GANEM B, 1993, CHEMTRACTS ORG CHEM, V6, P1
[9] MASS-SPECTROMETRIC DETECTION OF THE NONCOVALENT GDP-BOUND CONFORMATIONAL STATE OF THE HUMAN H-RAS PROTEIN
GANGULY, AK
PRAMANIK, BN
TSARBOPOULOS, A
COVEY, TR
HUANG, E
FUHRMAN, SA
[J]. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1992, 114 (16) : 6559 - 6560
[10] GASKELL SJ, 1986, MASS SPECTROMOETRY B

← 1 2 3 4 →