細胞内情報伝達研究における新技術ならびにツールの開発

細胞内情報伝達研究における新技術ならびにツールの開発

独自の解析法や研究ツールの開発に取り組んでいます

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更新日 2016-07-12 | 作成日 2008-01-20

細胞内情報伝達研究における新技術ならびにツールの開発

細胞内情報伝達研究における新技術ならびにツールの開発

研究には、様々な実験手法や研究用ツールが使われているが、これまでにも新しい技術の開発により、研究が飛躍的に進展したという例は数多く知られている。我々は、様々な生命現象とプロテインキナーゼの関わりを網羅的に解析することを目指して、プロテインキナーゼの活性検出法や発現パターン解析法などに取り組んできた。また、脱リン酸化酵素であるプロテインホスファターゼの解析法や特異的阻害剤の探索だけでなく、タンパク質リン酸化に関連した様々な解析法の開発を手がけている。

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発表論文

(1)日本語総説

杉山康憲、片山将一、馬場裕美、末吉紀行、亀下勇
液相等電点泳動を用いた二次元泳動法の活用
電気泳動 58, 62-64 (2014)

石田敦彦、亀下勇
インゲルキナーゼアッセイ法 〜その後の進歩
電気泳動 58, 62-64 (2014)

亀下勇、末吉紀行
プロテインキナーゼ解析用ツールとしての網羅的抗体
生化学 83, 330-335 (2011)

茂里康、石田敦彦、亀下 勇
“プロテインキナーゼに関する最近の検出技術—より網羅的に、よりハイスループットに”
化学と生物 45, 252-260 (2007)

(2)学術論文

Senga, Y.*, Akizuki, K.*, Katayama, S., Shigeri, Y., Kameshita, I., Ishida, A., Sueyoshi, N.
"High-performance CaMKI: A highly active and stable form of CaMKIδ produced by high-level soluble expression in Escherichia coli."
Biochem. Biophys. Res. Commun. 475, 277-282 (2016) *: The first two authors contributed equally to this work.
この論文の日本語ダイジェストはこちら

Sugiyama, Y., Katayama, S., Kameshita, I., Morisawa, K., Higuchi, T., Todaka, H., Kinoshita, E., Kinoshita-Kikuta, E., Koike, T., Taniguchi, T., Sakamoto, S.
"Expression and phosphorylation state analysis of intracellular protein kinases using Multi-PK antibody and Phos-tag SDS-PAGE."
MethodsX 2, 469-474 (2015)

Kameshita, I., Yamashita, S., Katayama, S., Senga, Y., Sueyoshi, N.
"TandeMBP: generation of a unique protein substrate for protein kinase assays."
J. Biochem. 156, 147-154 (2014)

Kaneko, K., Sueyoshi, N., Kameshita, I., Ishida, A.
"Ink-native electrophoresis: an alternative to blue-native electrophoresis more suitable for in-gel detection of enzymatic activity."
Anal. Biochem. 440, 142-144 (2013)

Baba, H., Masuda, Y., Sueyoshi, N., Kameshita, I.
"In gel phosphatase assay using non-denaturing two-dimensional electrophoresis."
J. Biochem. 152, 557-563 (2012)

Senga, Y., Nagamine, T., Sekiguchi, M., Kaneko, K., Sueyoshi, N., Kameshita, I.
"Detection of protein kinase substrates in tissue extracts after separation by isoelectric focusing."
Anal. Biochem. 408, 345-347 (2011)

Ishida, A., Kameshita, I.
"In-gel protein phosphatase assays and other useful methods for the detection of protein phosphatase activities."
Anticancer Agents Med. Chem. 11, 47-53 (2011)

Sugiyama, Y., Murao, K., Imachi, H., Sueyoshi, N., Ishida, T., Kameshita, I.
"Calcium/calmodulin-dependent protein kinase IV involvement in the pathophysiology of glucotoxicity in rat pancreatic beta-cells."
Metabolism. 60, 145-153 (2011)

Kameshita, I., Baba, H., Umeda, Y., Sueyoshi, N.
"In-gel protein phosphatase assay using fluorogenic substrates."
Anal. Biochem. 400, 118-122 (2010)

Sugiyama, Y., Hatano, N., Sueyoshi, N., Suetake, I., Tajima, S., Kinoshita, E., Kinoshita-Kikuta, E., Koike, T., Kameshita, I.
"The DNA-binding activity of mouse DNA methyltransferase 1 is regulated by phosphorylation with casein kinase 1delta/epsilon."
Biochem. J. 427, 489-497 (2010)

Sugiyama, Y., Ishida, A., Sueyoshi, N., Kameshita, I.
"Tyrosine kinase activity of a Ca2+/calmodulin-dependent protein kinase II catalytic fragment. "
Biochem. Biophys. Res. Commun. 377, 648-652 (2008)

Sugiyama, Y., Shimomura, S., Sueyoshi, N., Kameshita, I. :
“Two-dimensional gel electrophoretic analysis of CNBr fragments containing subdomain VIB of protein kinases using Multi-PK antibody”
Anal. Biochem. 373, 173-175 (2008)

Sueyoshi, N., Takao, T., Nimura, T., Sugiyama, Y., Numano, T., Shigeri, Y., Taniguchi, T., Kameshita, I., Ishida, A. :
“Inhibitors of the Ca2+/calmodulin-dependent protein kinase phosphatase family (CaMKP and CaMKP-N)”
Biochem. Biophys. Res. Commun. 363, 715-721 (2007)

Sugiyama, Y., Sueyoshi, N., Kameshita, I.
“Two-dimensional expression pattern analysis of protein kinases after separation by MicroRotofor/SDS-PAGE”
Anal. Biochem. 359, 271-273 (2006)

Shoju, H., Sueyoshi, N., Kameshita, I.
“High-level expression of proteins in Escherichia coli using pETCX10 expression system”
Anal. Biochem. 353, 290-293 (2006)

Sugiyama, Y., Sueyoshi, N., Shigeri, Y., Tatsu, Y., Yumoto, N., Ishida, A., Taniguchi, T., Kameshita, I.
“Generation and application of a monoclonal antibody that detect a wide variety of protein tyrosine kinases”
Anal. Biochem. 347, 112-120 (2005)

Shoju, H., Sueyoshi, N., Ishida, A., Kameshita, I.
“High level expression and preparation of autonomous Ca2+/calmodulin- dependent protein kinase II in Escherichia coli”
J. Biochem. 138, 605-611 (2005)

Kameshita, I., Kinoshita, S., Shigeri, Y., Tatsu, Y., Yumoto, N., Ishida, A.
“Generation of a polyclonal antibody that simultaneously detects multiple Ser/Thr protein kinases”
J. Biochem. Biophys. Methods 60, 13-22 (2004)

Kameshita, I., Tsuge, T., Kinashi, T., Kinoshita, S., Sueyoshi, N., Ishida, A., Taketani, S., Shigeri, Y., Tatsu, Y., Yumoto, N., Okazaki, K.
“A new approach for the detection of multiple protein kinases using monoclonal antibodies directed to the highly conserved region of protein kinases”
Anal. Biochem. 322, 215-224 (2003)

Ishida, A., Kameshita, I., Okuno, S., Kitani, T., Fujisawa, H.
“Phosphorylation of Calmodulin by Ca2+/Calmodulin-dependent Protein Kinase IV”
Arch. Biochem. Biophys. 407, 72-82 (2002)

Kameshita, I., Taketani, S., Ishida, A., Fujisawa, H.
“Detection of a variety of Ser/Thr protein kinases using a synthetic peptide with multiple phosphorylation sites”
J. Biochem. 126, 991-995 (1999)

Tatsu, Y. Shigeri, Y., Ishida, A., Kameshita, I., Fujisawa, H., Yumoto, N.
“Synthesis of caged peptides using caged lysine: application to the synthesis of caged AIP, a highly specific inhibitor of calmodulin-dependent protein kinase II”
Bioorg. Med. Chem. Lett. 9, 1093-1096 (1999)

Kameshita, I., Ishida, A., Fujisawa, H.
“Analysis of protein-protein interaction by two-dimensional affinity electrophoresis”
Anal. Biochem. 262, 90-92 (1998)

Ishida, A., Shigeri, Y., Tatsu, Y., Uegaki, K., Kameshita, I., Okuno, S., Kitani, T., Yumoto, N., Fujisawa, H.
"Critical amino acid residues of AIP, a highly specific inhibitory peptide of calmodulin-dependent protein kinase II"
FEBS Lett. 427, 115-118 (1998)

Ishida, A., Kameshita, I., Fujisawa, H.
“Assay of protein phosphatase activities with phosphopeptide-magnetic particle conjugates”
Anal. Biochem. 254, 152-155 (1997)

Kameshita, I., Ishida, A., Fujisawa, H.
“Detection of protein phosphatase activities in sodium dodecyl sulfate-polyacrylamide gel using peptide substrates”
Anal. Biochem. 245, 149-153 (1997)

Kameshita, I., Ishida, A., Fujisawa, H.
”New peptide conjugate as a highly specific substrate for MAP kinase”
J. Biochem. 122, 168-172 (1997)

Kameshita, I., Fujisawa, H.
“Detection of calcium binding proteins by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis”
Anal. Biochem. 249, 252-255 (1997)

Kameshita, I., Fujisawa, H.
“Detection of protein kinase activities toward oligopeptides in sodium sodecyl sulfate-polyacrylamide gel”
Anal. Biochem. 237, 198-203 (1996)

Ishida, A., Kameshita, I., Okuno, S., Kitani, T., Fujisawa, H.
“A novel highly specific and potent inhibitor of calcium-dependent protein kinase II”
Biochem. Biophys. Res. Commun. 212, 806-812 (1995)