Direct Quantification of Protein-Metal Ion Affinities by Electrospray Ionization Mass Spectrometry

被引：32
作者：
Deng, Lu
Sun, Nian
Kitova, Elena N.
Klassen, John S. [1 ]
机构：
[1] Univ Alberta, Alberta Ingenu Ctr Carbohydrate Sci, Edmonton, AB T6G 2G2, Canada
来源：
ANALYTICAL CHEMISTRY | 2010年 / 82卷 / 06期
基金：
加拿大自然科学与工程研究理事会;
关键词：
NANOELECTROSPRAY IONIZATION; BETA-LACTOGLOBULIN; MODEL SYSTEM; BINDING; COMPLEXES; CALCIUM; CONSTANTS; ZINC; ESI;
D O I：
10.1021/ac902633d
中图分类号：
O65 [分析化学];
学科分类号：
070302 ; 081704 ;
摘要：
The application of the direct electrospray ionization mass spectrometry (ES-MS) assay for quantifying the stoichiometry and absolute affinity of protein-metal ion binding in vitro is described. Control ES-MS experiments performed on solutions containing calcium chloride or calcium acetate and a pair of proteins that do not bind calcium ions in solution revealed that the nonspecific association of metal ions to proteins during ES is a random process, independent of protein size and structure. These results establish the reliability of the reference protein method for quantitatively correcting ES mass spectra for the occurrence of nonspecific metal ion binding to proteins during ES-MS analysis. To demonstrate the utility of the direct ES-MS assay, when carried out using the reference protein method, the calcium binding stoichiometry of bovine alpha-lactalbumin and the calcium ion affinity of bovine beta-lactoglobulin were established.
引用
收藏
页码：2170 / 2174
页数：5
相关论文
共 29 条
[1]   STRUCTURE OF HUMAN LACTOFERRIN AT 3.2-A RESOLUTION [J].
ANDERSON, BF ;
BAKER, HM ;
DODSON, EJ ;
NORRIS, GE ;
RUMBALL, SV ;
WATERS, JM ;
BAKER, EN .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1987, 84 (07) :1769-1773
[2]  
BERG JM, 1990, ANNU REV BIOPHYS BIO, V19, P405
[3]   Quantitative determination of noncovalent binding interactions using soft ionization mass spectrometry [J].
Daniel, JM ;
Friess, SD ;
Rajagopalan, S ;
Wendt, S ;
Zenobi, R .
INTERNATIONAL JOURNAL OF MASS SPECTROMETRY, 2002, 216 (01) :1-27
[4]   BETA-LACTOGLOBULIN AND ALPHA-LACTALBUMIN AS POTENTIAL MODULATORS OF MAMMARY CELLULAR-ACTIVITY - A CA2+-RESPONSIVE MODEL SYSTEM USING ACID PHOSPHOPROTEIN PHOSPHATASES [J].
FARRELL, HM ;
THOMPSON, MP .
PROTOPLASMA, 1990, 159 (2-3) :157-167
[5]   Structural changes in calcium-binding allergens: use of circular dichroism to study binding characteristics [J].
Hebenstreit, D ;
Ferreira, F .
ALLERGY, 2005, 60 (09) :1208-1211
[6]   Investigation of intact protein complexes by mass spectrometry [J].
Heck, AJR ;
van den Heuvel, RHH .
MASS SPECTROMETRY REVIEWS, 2004, 23 (05) :368-389
[7]   The bundlin pilin protein of enteropathogenic Escherichia coli is an N-acetyllactosamine-specific lectin [J].
Hyland, Romney M. ;
Sun, Jiangxiao ;
Griener, Thomas P. ;
Mullvey, George L. ;
Klassen, John S. ;
Donnenberg, Michael S. ;
Armstrong, Glen D. .
CELLULAR MICROBIOLOGY, 2008, 10 (01) :177-187
[8]   Which electrospray-based ionization method best reflects protein-ligand interactions found in solution? A comparison of ESI, nanoESI, and ESSI for the determination of dissociation constants with mass spectrometry [J].
Jecklin, Matthias Conradin ;
Touboul, David ;
Bovet, Cedric ;
Wortmann, Arno ;
Zenobi, Renato .
JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY, 2008, 19 (03) :332-343
[9]   Investigation of structure, dynamics and function of metalloproteins with electrospray ionization mass spectrometry [J].
Kaltashov, Igor A. ;
Zhang, Mingxuan ;
Eyles, Stephen J. ;
Abzalimov, Rinat R. .
ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 2006, 386 (03) :472-481
[10]   Affinities of Shiga toxins 1 and 2 for univalent and oligovalent Pk-trisaccharide analogs measured by electrospray ionization mass spectrometry [J].
Kitova, Elena N. ;
Kitov, Pavel I. ;
PaszkiewiCZ, Eugenia ;
Kim, Jonghwa ;
Mulvey, George L. ;
Armstrong, Glen D. ;
Bundle, David R. ;
Klassen, John S. .
GLYCOBIOLOGY, 2007, 17 (10) :1127-1137
123