A two-dimensional electrophoresis map of Chinese hamster ovary cell proteins based on fluorescence staining

被引：43

作者：

Hayduk, EJ ^{[1
]}

Choe, LH ^{[1
]}

Lee, KH ^{[1
]}

机构：

[1] Cornell Univ, Sch Chem & Biomol Engn, Ithaca, NY 14850 USA

来源：

ELECTROPHORESIS | 2004年 / 25卷 / 15期

关键词：

Chinese hamster ovary cells; pro-Q diamond; proteomics; SYPRO Ruby; tandem time-of-flight;

D O I：

10.1002/elps.200406010

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We report on the development of a detailed two-dimensional electrophoresis map of Chinese hamster ovary (CHO) cell proteins based on fluorescence staining and tandem time-of-flight (TOF/TOF)-mass spectrometry. We observed a 71% success rate in the identification of proteins even though the CHO genome is not sequenced. The map consists of 224 protein identifications present in 274 two-dimensional gel electrophoresis (2-DE) gel spots. We have also initiated a study of the phosphoproteome using a commercially available phosphoprotein-specific fluorescent stain. Using this stain, we observe 672 phosphorylated proteins, including many proteins known to be phosphorylated, which is 36% of the proteins we visualized with a total protein stain and consistent with expectations.

引用

页码：2545 / 2556

页数：12

共 32 条

[1] Regulation of actin dynamics through phosphorylation of cofilin by LIM-kinase [J].

Arber, S ;

Barbayannis, FA ;

Hanser, H ;

Schneider, C ;

Stanyon, CA ;

Bernard, O ;

Caroni, P .

NATURE, 1998, 393 (6687) :805-809

[2] Cloning and characterization of the p42 subunit of mammalian translation initiation factor 3 (eIF3): demonstration that eIF3 interacts with eIF5 in mammalian cells [J].

Bandyopadhyay, A ;

Maitra, U .

NUCLEIC ACIDS RESEARCH, 1999, 27 (05) :1331-1337

[3] Regulation of peptide-chain elongation in mammalian cells [J].

Browne, GJ ;

Proud, CG .

EUROPEAN JOURNAL OF BIOCHEMISTRY, 2002, 269 (22) :5360-5368

[4]

Champion KM, 1999, ELECTROPHORESIS, V20, P994, DOI 10.1002/(SICI)1522-2683(19990101)20:4/5<994::AID-ELPS994>3.0.CO

[5]

2-N

[6] THE MAJOR PHOSPHORYLATION SITE OF NUCLEOPHOSMIN (B23) IS PHOSPHORYLATED BY A NUCLEAR KINASE-II [J].

CHAN, PK ;

LIU, QR ;

DURBAN, E .

BIOCHEMICAL JOURNAL, 1990, 270 (02) :549-552

[7] Quantitative and qualitative measure of intralaboratory two-dimensional protein gel reproducibility and the effects of sample preparation, sample load, and image analysis [J].

Choe, LH ;

Lee, KH .

ELECTROPHORESIS, 2003, 24 (19-20) :3500-3507

[8]

Choe LH, 1999, ELECTROPHORESIS, V20, P798, DOI 10.1002/(SICI)1522-2683(19990101)20:4/5<798::AID-ELPS798>3.0.CO

[9]

2-F

[10]

Constantin B, 1998, J CELL SCI, V111, P1695

← 1 2 3 4 →