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       Journal List > Exp Mol Med > v.40(4); Aug 2008
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Exp Mol Med. 2008 Aug;40(4):377-386.
Published online 2008 August 31.  doi: 10.3858/emm.2008.40.4.377.
Copyright © 2008 Korean Society of Medical Biochemistry and Molecular Biology
Transient phosphorylation of tumor associated microtubule associated protein (TMAP)/cytoskeleton associated protein 2 (CKAP2) at Thr-596 during early phases of mitosis
Kyung Uk Hong,1 Yong-Bock Choi,2 Jung-Hwa Lee,2 Hyun-Jun Kim,1 Hye-Rim Kwon,1 Yeon-Sun Seong,3 Heung Tae Kim,2 Joobae Park,1 Chang-Dae Bae,1 and Kyeong-Man Hong2
1Department of Molecular Cell Biology and Samsung Biomedical Research Institute, Sungkyunkwan University School of Medicine, Suwon 440-769, Korea.
2Research Institute, National Cancer Center, Goyang 410-769, Korea.
3Department of Biochemistry, College of Medicine, Dankook University, ChunAn 330-714, Korea.

Corresponding author: Tel, 82-31-920-2261; Fax, 82-31-920-2006; Email: kmhong@ncc.re.kr (K.M. Hong); Tel, 82-31-299-6130; Fax, 82-31-299-6149; Email: cdbae@med.skku.ac.kr (C.D. Bae)
Accepted April 07, 2008.

Abstract

Tumor associated microtubule associated protein (TMAP), also known as cytoskeleton associated protein 2 (CKAP2) is a mitotic spindle-associated protein whose expression is cell cycle-regulated and also frequently deregulated in cancer cells. Two monoclonal antibodies (mAbs) against TMAP/CKAP2 were produced: B-1-13 and D-12-3. Interestingly, the reactivity of mAb D-12-3 to TMAP/CKAP2 was markedly decreased specifically in mitotic cell lysate. The epitope mapping study showed that mAb D-12-3 recognizes the amino acid sequence between 569 and 625 and that phosphorylation at T596 completely abolishes the reactivity of the antibody, suggesting that the differential reactivity originates from the phosphorylation status at T596. Immunofluorescence staining showed that mAb D-12-3 fails to detect TMAP/CKAP2 in mitotic cells between prophase and metaphase, but the staining becomes evident again in anaphase, suggesting that phosphorylation at T596 occurs transiently during early phases of mitosis. These results suggest that the cellular functions of TMAP/CKAP2 might be regulated by timely phosphorylation and dephosphorylation during the course of mitosis.

Keywords: antibodies, monoclonal, cell cycle, CKAP2 protein, human, fluorescent antibody technique, direct, phosphorylation.
Abbreviations
CKAP2 cytoskeleton associated protein 2
TMAP tumor associated microtubule associated protein
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