期刊
EXPERIMENTAL AND MOLECULAR MEDICINE
卷 40, 期 4, 页码 377-386出版社
NATURE PUBLISHING GROUP
DOI: 10.3858/emm.2008.40.4.377
关键词
antibodies, monoclonal; cell cycle; CKAP2 protein, human; fluorescent antibody technique, direct; phosphorylation
资金
- National Cancer Center, Korea [0510370, 0810240]
- National R&D Program for Cancer Control, Ministry of Health and Welfare, Republic of Korea [0720370]
- Molecular and Cellular Bio-Discovery Project, KISTEP [2004-01789]
- Korea Health Promotion Institute [0720370] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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.
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