Document Detail

Anti-phosphopeptide antibody, P-STM as a novel tool for detecting mitotic phosphoproteins: identification of lamins A and C as two major targets.
MedLine Citation:
PMID:  15597429     Owner:  NLM     Status:  MEDLINE    
A polyclonal, phospho-epitope-specific antibody (P-STM) was generated to detect the activated p21-activated kinase 2 (PAK2), based on the regulatory autophosphorylation site Thr(402) of PAK2 [Yu et al., 1998]. In this report, we show that this antibody can also recognize many phosphoproteins in mitotic HeLa and A431 cells. Signal of these phosphoproteins emerged after treating the cells with nocodazole and okadaic acid, and was highly detected in G2-M phase transition of HeLa cells released from double thymidine block. Immunofluorescence analysis revealed that P-STM strongly stained HeLa cells at prometaphase and metaphase, but not at interphase and anaphase. Interestingly, this staining pattern was almost identical to that obtained by staining with MPM2, a monoclonal antibody known to react with phosphoproteins in mitotic HeLa cells. However, the phosphoproteins detected by the two antibodies are quite different. Two-dimensional gel electrophoresis (2DE) and tryptic peptide fingerprint analysis by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry were employed to identify lamins A and C as two of the mitotic cell-specific phosphoproteins recognized by P-STM. Lamins A and C immunoprecipitated from nocodazole-treated cells, but not from untreated cells showed strong reactivity to P-STM, and this reactivity lost completely after protein phosphatase 2A treatment. In summary, our results show that P-STM represents a novel tool for detecting mitotic phosphoproteins, which are different from those recognized by MPM2, and that lamins A and C are the two prominent mitotic phosphoproteins detected by P-STM.
I-Chen Tsai; Ya-Ju Hsieh; Ping-Chiang Lyu; Jau-Song Yu
Related Documents :
22552949 - N-glycan biosynthesis inhibitors induce in vitro anticancer activity in colorectal canc...
9136899 - Icrf-193, a catalytic inhibitor of dna topoisomerase ii, delays the cell cycle progress...
6879589 - Unique morphology of hela cell attachment, spreading and detachment from microcarrier b...
1280979 - Selenite-induced inhibition of colony formation by buthionine sulfoximine-sensitive and...
17825929 - Comparison of hela-i, hep-2 and nci-h292 cell lines for the isolation of human respirat...
1315289 - The 180-kda isoform of topoisomerase ii is localized in the nucleolus and belongs to th...
15601319 - Cell cycle arrest of human gingival fibroblasts and periodontal ligament cells by actin...
6406519 - Formation of mast cell granules in cell cycle mutants of an undifferentiated mastocytom...
2200449 - Neurite outgrowth in response to transfected n-cam changes during development and is mo...
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of cellular biochemistry     Volume:  94     ISSN:  0730-2312     ISO Abbreviation:  J. Cell. Biochem.     Publication Date:  2005 Apr 
Date Detail:
Created Date:  2005-03-15     Completed Date:  2005-09-26     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8205768     Medline TA:  J Cell Biochem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  967-81     Citation Subset:  IM    
Copyright Information:
(c) 2005 Wiley-Liss, Inc.
Department of Cell and Molecular Biology, Graduate Institute of Basic Medical Sciences, Medical College of Chang Gung University, Tao-Yuan, Taiwan, Republic of China.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Amino Acid Sequence
Antibodies / immunology*
Electrophoresis, Gel, Two-Dimensional
Hela Cells
Lamin Type A / chemistry,  metabolism*
Molecular Sequence Data
Phosphopeptides / immunology*
Phosphoproteins / analysis*,  immunology
Sequence Homology, Amino Acid
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Reg. No./Substance:
0/Antibodies; 0/Lamin Type A; 0/Phosphopeptides; 0/Phosphoproteins; 0/lamin C

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

Previous Document:  System zones in capillary zone electrophoresis.
Next Document:  A blended infant massage--parenting enhancement program for recovering substance-abusing mothers.