Characterization of Tetrahymena histone H2B variants and posttranslational populations by electron capture dissociation (ECD) Fourier transform ion cyclotron mass spectrometry (FT-ICR MS)

被引:81
作者
Medzihradszky, KF
Zhang, X
Chalkley, RJ
Guan, S
McFarland, MA
Chalmers, MJ
Marshall, AG
Diaz, RL
Allis, CD
Burlingame, AL
机构
[1] Univ Calif San Francisco, Dept Pharmaceut Chem, San Francisco, CA 94143 USA
[2] Univ Calif San Francisco, Mass Spectrometry Facil, San Francisco, CA 94143 USA
[3] Natl High Magnet Field Lab, Ion Cyclotron Resonance Program, Tallahassee, FL 32310 USA
[4] Rockefeller Univ, Lab Chromatin Biol, New York, NY 10021 USA
关键词
D O I
10.1074/mcp.M400041-MCP200
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
This work describes the nature and sequence information content of the electron capture dissociation mass spectra for the intact Tetrahymena histone H2B. Two major variants of this protein were present bearing nominal modifications of both + 42 and + 84 Da. This work describes identification of the nature of these two modifications. For example, using gas-phase selection and isolation of the + 42-Da modified species, from a background of two H2B variants each present in six or more posttranslationally modified isoforms, we were able to determine that this + 42-Da modification isoform bears trimethylation rather than acetylation. LC-CIDMS analysis was also employed on digested preparations to obtain complementary detail of the nature of site-specific posttranslational modifications. This study establishes that integration of the information from these two datasets provides a comprehensive map of posttranslational occupancy for each particular covalent assemblage selected for structural investigation.
引用
收藏
页码:872 / 886
页数:15
相关论文
共 47 条
[21]   Localization of labile posttranslational modifications by electron capture dissociation:: The case of γ-carboxyglutamic acid [J].
Kelleher, RL ;
Zubarev, RA ;
Bush, K ;
Furie, B ;
Furie, BC ;
McLafferty, FW ;
Walsh, CT .
ANALYTICAL CHEMISTRY, 1999, 71 (19) :4250-4253
[22]   Complete characterization of posttranslational modification sites in the bovine milk protein PP3 by tandem mass spectrometry with electron capture dissociation as the last stage [J].
Kjeldsen, F ;
Haselmann, KF ;
Budnik, BA ;
Sorensen, ES ;
Zubarev, RA .
ANALYTICAL CHEMISTRY, 2003, 75 (10) :2355-2361
[23]   APPLICATION OF ELECTROSPRAY AND FAST-ATOM-BOMBARDMENT MASS-SPECTROMETRY TO THE IDENTIFICATION OF POSTTRANSLATIONAL AND OTHER CHEMICAL MODIFICATIONS OF PROTEINS AND PEPTIDES [J].
KOUACH, M ;
BELAICHE, D ;
JAQUINOD, M ;
COUPPEZ, M ;
KMIECIK, D ;
RICART, G ;
VANDORSSELAER, A ;
SAUTIERE, P ;
BRIAND, G .
BIOLOGICAL MASS SPECTROMETRY, 1994, 23 (05) :283-294
[24]   Analysis of a piwi-related gene implicates small RNAs in genome rearrangement in tetrahymena [J].
Mochizuki, K ;
Fine, NA ;
Fujisawa, T ;
Gorovsky, MA .
CELL, 2002, 110 (06) :689-699
[25]   N-TRIMETHYLALANINE, A NOVEL BLOCKED N-TERMINAL RESIDUE OF TETRAHYMENA HISTONE-H2B [J].
NOMOTO, M ;
KYOGOKU, Y ;
IWAI, K .
JOURNAL OF BIOCHEMISTRY, 1982, 92 (05) :1675-1678
[26]   CHARACTERIZATION OF 2 TYPES OF HISTONE H2B GENES FROM MACRONUCLEI OF TETRAHYMENA-THERMOPHILA [J].
NOMOTO, M ;
IMAI, N ;
SAIGA, H ;
MATSUI, T ;
MITA, T .
NUCLEIC ACIDS RESEARCH, 1987, 15 (14) :5681-5697
[27]   Assembly of cell regulatory systems through protein interaction domains [J].
Pawson, T ;
Nash, P .
SCIENCE, 2003, 300 (5618) :445-452
[28]   Interaction domains: from simple binding events to complex cellular behavior [J].
Pawson, T ;
Raina, M ;
Nash, P .
FEBS LETTERS, 2002, 513 (01) :2-10
[29]   Processive phosphorylation of p130Cas by Src depends on SH3-polyproline interactions [J].
Pellicena, P ;
Miller, WT .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (30) :28190-28196
[30]   Shotgun annotation of histone modifications: A new approach for streamlined characterization of proteins by top down mass spectrometry [J].
Pesavento, JJ ;
Kim, YB ;
Taylor, GK ;
Kelleher, NL .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2004, 126 (11) :3386-3387