| RRC ID |
1700
|
| 著者 |
Uchiyama S, Kobayashi S, Takata H, Ishihara T, Sone T, Matsunaga S, Fukui K.
|
| タイトル |
Protein composition of human metaphase chromosomes analyzed by two-dimensional electrophoreses.
|
| ジャーナル |
Cytogenet Genome Res
|
| Abstract |
A large amount of metaphase chromosomes were isolated from synchronized human cell lines by a polyamine procedure. All the chromosomal proteins extracted by an acetic acid extraction method were fully dissolved into the sample solutions for isoelectric focusing (IEF) or radical free and highly reduced (RFHR) two-dimensional electrophoreses (2-DEs). As a result, well-separated and highly reproducible 2-DE patterns were obtained. This could not be attained by an ordinary acetone precipitation method. The 2-DE patterns visualized using Coomassie Brilliant Blue (CBB) staining indicated that more than one hundred proteins were involved in the isolated metaphase chromosomes, although the most abundant proteins, histones, occupied a greater part of the chromosomal proteins. It was also shown that colcemid treatment for cell cycle synchronization had little effect on the 2-DE pattern compared to that obtained without the treatment. Furthermore, no significant differences were observed in the 2-DE patterns among the chromosomal proteins prepared from two different human cell lines, BALL-1 and K562. However, 2-DE analysis of isolated metaphase chromosomes from HeLa cells apparently showed a smaller number of proteins than the BALL-1 and K562 cell lines at a neutral pI range. The present study paves the way for elucidating protein composition of human metaphase chromosomes.
|
| 巻・号 |
107(1-2)
|
| ページ |
49-54
|
| 公開日 |
2004-1-1
|
| DOI |
10.1159/000079571
|
| PII |
79571
|
| PMID |
15305056
|
| MeSH |
Cell Line, Tumor
Chromosomes, Human / chemistry*
Chromosomes, Human / genetics*
Electrophoresis, Gel, Two-Dimensional / methods*
HeLa Cells / chemistry
HeLa Cells / metabolism
Histones / metabolism
Humans
Isoelectric Focusing / methods
K562 Cells / chemistry
K562 Cells / metabolism
Metaphase / genetics*
Nucleoproteins / metabolism*
Peptide Mapping / methods
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods
|
| IF |
1.114
|
| 引用数 |
11
|
|
WOS 分野
|
GENETICS & HEREDITY
CELL BIOLOGY
|
| リソース情報 |
| 情報 |
|
| ヒト・動物細胞 |
BALL-1(RCB0256)
K562(RCB0027) |
