Supercritical fluid chromatography reduction of hydrogen/deuterium back exchange in solution-phase hydrogen/deuterium exchange with mass spectrometric analysis

被引：28

作者：

Emmett, Mark R.

Kazazic, Sasa

Marshall, Alan G.

Chen, Wei

Shi, Stone D. -H.

Bolanos, Ben

Greig, Michael J.

机构：

[1] Florida State Univ, Natl High Magnet Field Lab, Ion Cyclotron Resonance Program, Tallahassee, FL 32310 USA

[2] Florida State Univ, Dept Chem & Biochem, Tallahassee, FL 32306 USA

[3] Pfizer Inc, Global Res & Dev, La Jolla Labs, San Diego, CA 92121 USA

来源：

ANALYTICAL CHEMISTRY | 2006年 / 78卷 / 19期

关键词：

PROTEIN-PROTEIN INTERACTIONS; CHEMICAL CROSS-LINKING; IDENTIFICATION; PEPTIDES; SURFACES;

D O I：

10.1021/ac060693n

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

The single biggest problem with solution-phase H/D exchange as a mass spectrometric probe of surface exposure in a protein (or protein complex) is back-exchange of H for D after the initial H/D exchange has been quenched. Back-exchange results in loss of pertinent data and also greatly hampers data analysis. Previously, very fast, cold (0-4 degrees C) HPLC was performed to help reduce back-exchange, but calculated back-exchange still averages similar to 30%. In this report, supercritical fluid chromatography replaces HPLC as the desalting/separation technique prior to mass analysis, providing a dramatic reduction in back-exchange compared to the fast, cold HPLC methods.

引用

页码：7058 / 7060

页数：3

共 20 条

[1]

Blackwell JA, 1999, HRC-J HIGH RES CHROM, V22, P74

[2] SFC/MS in drug discovery at Pfizer, La Jolla [J].

Bolaños, B ;

Greig, M ;

Ventura, M ;

Farrell, W ;

Aurigemma, CM ;

Li, H ;

Quenzer, TL ;

Tivel, K ;

Bylund, JMR ;

Tran, P ;

Pham, C ;

Phillipson, D .

INTERNATIONAL JOURNAL OF MASS SPECTROMETRY, 2004, 238 (02) :85-97

[3] Chemistry for the analysis of protein-protein interactions: Rapid and efficient cross-linking triggered by long wavelength light [J].

Fancy, DA ;

Kodadek, T .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (11) :6020-6024

[4] Identification of the sites of hydroxyl radical reaction with peptides by hydrogen/deuterium exchange: Prevalence of reactions with the side chains [J].

Goshe, MB ;

Chen, YH ;

Anderson, VE .

BIOCHEMISTRY, 2000, 39 (07) :1761-1770

[5]

Greig M.J., 2001, 49 ASMS C MASS SPECT

[6] Mapping the ribosomal RNA neighborhood of protein L11 by directed hydroxyl radical probing [J].

Holmberg, L ;

Noller, HF .

JOURNAL OF MOLECULAR BIOLOGY, 1999, 289 (02) :223-233

[7] Automated extraction of backbone deuteration levels from amide H/2 H mass spectrometry experiments [J].

Hotchko, M ;

Anand, GS ;

Komives, EA ;

Ten Eyck, LF .

PROTEIN SCIENCE, 2006, 15 (03) :583-601

[8] CHEMICAL IONIZATION MASS-SPECTRA OF HIGH MOLECULAR-WEIGHT, BIOLOGICALLY-ACTIVE COMPOUNDS PRODUCED FOLLOWING SUPERCRITICAL FLUID CHROMATOGRAPHY [J].

KALINOSKI, HT ;

WRIGHT, BW ;

SMITH, RD .

BIOMEDICAL AND ENVIRONMENTAL MASS SPECTROMETRY, 1988, 15 (04) :239-242

[9] Chemical cross-linking and high-performance Fourier transform ion cyclotron resonance mass spectrometry for protein interaction analysis: Application to a calmodulin/target peptide complex [J].

Kalkhof, S ;

Ihling, C ;

Mechtler, K ;

Sinz, A .

ANALYTICAL CHEMISTRY, 2005, 77 (02) :495-503

[10] Identification of subunit-subunit interactions in bacteriophage P22 procapsids by chemical cross-linking and mass spectrometry [J].

Kang, S ;

Hawkridge, AM ;

Johnson, KL ;

Muddiman, DC ;

Prevelige, PE .

JOURNAL OF PROTEOME RESEARCH, 2006, 5 (02) :370-377

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