CTGF/Hcs24 interacts with the cytoskeletal protein actin in chondrocytes

Gen Yosimichi; Satoshi Kubota; Takako Hattori; Takashi Nishida; Kumiko Nawachi; Tohru Nakanishi; Masafumi Kamada; Teruko Takano-Yamamoto; Masaharu Takigawa

doi:10.1016/S0006-291X(02)02739-0

CTGF/Hcs24 interacts with the cytoskeletal protein actin in chondrocytes

Gen Yosimichi, Satoshi Kubota, Takako Hattori, Takashi Nishida, Kumiko Nawachi, Tohru Nakanishi, Masafumi Kamada, Teruko Takano-Yamamoto, Masaharu Takigawa

Research output: Contribution to journal › Article › peer-review

9 Citations (Scopus)

Abstract

Connective tissue growth factor/hypertrophic chondrocyte-specific gene product 24 (CTGF/Hcs24) displays multiple functions in several types of mesenchymal cells, including the promotion of proliferation and differentiation of chondrocytes. Recently, the internalization and intracellular function of CTGF/Hcs24 were indicated as well. In this study, a binding protein for this factor was purified from the cytosolic fraction of human chondrosarcoma-derived chondrocytic cell line (HCS-2/8) by CTGF/Hcs24-affinity chromatography. The apparent molecular weight of the protein was 42 kDa and determination of the internal amino acid sequence revealed this protein to be β- or γ-actin. An in vitro competitive binding assay of ¹²⁵I-labeled recombinant CTGF/Hcs24 with cold-rCTGF/Hcs24 showed that the binding between actin and ¹²⁵I-CTGF/Hcs24 was specific. Immunoprecipitation analysis also showed that CTGF/Hcs24 bound to actin in HCS-2/8 cells. However, rCTGF/Hcs24 had no effects on the expression level of γ-actin mRNA or total actin protein. These findings suggest that a significant portion of intracellular CTGF/Hcs24 may regulate certain cell biological events in chondrocytes through the interaction with this particular cytoskeletal protein.

Original language	English
Pages (from-to)	755-761
Number of pages	7
Journal	Biochemical and Biophysical Research Communications
Volume	299
Issue number	5
DOIs	https://doi.org/10.1016/S0006-291X(02)02739-0
Publication status	Published - 2002

Keywords

Actin
Cell cycle
Chondrocyte
Connective tissue growth factor/hypertrophic chondrocyte-specific gene product
Cytoskeleton
Purification

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/S0006-291X(02)02739-0

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@article{489b0a4efc614ba78459f59320cafa88,

title = "CTGF/Hcs24 interacts with the cytoskeletal protein actin in chondrocytes",

abstract = "Connective tissue growth factor/hypertrophic chondrocyte-specific gene product 24 (CTGF/Hcs24) displays multiple functions in several types of mesenchymal cells, including the promotion of proliferation and differentiation of chondrocytes. Recently, the internalization and intracellular function of CTGF/Hcs24 were indicated as well. In this study, a binding protein for this factor was purified from the cytosolic fraction of human chondrosarcoma-derived chondrocytic cell line (HCS-2/8) by CTGF/Hcs24-affinity chromatography. The apparent molecular weight of the protein was 42 kDa and determination of the internal amino acid sequence revealed this protein to be β- or γ-actin. An in vitro competitive binding assay of 125I-labeled recombinant CTGF/Hcs24 with cold-rCTGF/Hcs24 showed that the binding between actin and 125I-CTGF/Hcs24 was specific. Immunoprecipitation analysis also showed that CTGF/Hcs24 bound to actin in HCS-2/8 cells. However, rCTGF/Hcs24 had no effects on the expression level of γ-actin mRNA or total actin protein. These findings suggest that a significant portion of intracellular CTGF/Hcs24 may regulate certain cell biological events in chondrocytes through the interaction with this particular cytoskeletal protein.",

keywords = "Actin, Cell cycle, Chondrocyte, Connective tissue growth factor/hypertrophic chondrocyte-specific gene product, Cytoskeleton, Purification",

author = "Gen Yosimichi and Satoshi Kubota and Takako Hattori and Takashi Nishida and Kumiko Nawachi and Tohru Nakanishi and Masafumi Kamada and Teruko Takano-Yamamoto and Masaharu Takigawa",

note = "Funding Information: This work was supported in part by Grant-in-Aid for Science Research from the Research from the Ministry of Education, Science, Sports and Culture of Japan (S.K., M.T.); grants from the Research for the Future Programme of the Japan Society for the Promotion of Science (Biological Tissue Engineering Project, Contract Grant No. JSPS-RFTF98100201) (M.T.); the Naito Foundation (M.T.); the Nakatomi Health Science Foundation (M.T.); the Foundation for Growth Science in Japan (M.T.); and the Sumitomo Foundation (M.T.). We thank Dr. Terry D. Copeland for protein sequencing, Drs. Takanori Eguchi and Eiji Nakata for useful discussions, Drs. Masanao Minato and Norifumi Moritani for technical assistance, and Miss Yuki Nonami and Ayako Shizuku for secretarial assistance.",

year = "2002",

doi = "10.1016/S0006-291X(02)02739-0",

language = "English",

volume = "299",

pages = "755--761",

journal = "Biochemical and Biophysical Research Communications",

issn = "0006-291X",

publisher = "Academic Press Inc.",

number = "5",

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TY - JOUR

T1 - CTGF/Hcs24 interacts with the cytoskeletal protein actin in chondrocytes

AU - Yosimichi, Gen

AU - Kubota, Satoshi

AU - Hattori, Takako

AU - Nishida, Takashi

AU - Nawachi, Kumiko

AU - Nakanishi, Tohru

AU - Kamada, Masafumi

AU - Takano-Yamamoto, Teruko

AU - Takigawa, Masaharu

N1 - Funding Information: This work was supported in part by Grant-in-Aid for Science Research from the Research from the Ministry of Education, Science, Sports and Culture of Japan (S.K., M.T.); grants from the Research for the Future Programme of the Japan Society for the Promotion of Science (Biological Tissue Engineering Project, Contract Grant No. JSPS-RFTF98100201) (M.T.); the Naito Foundation (M.T.); the Nakatomi Health Science Foundation (M.T.); the Foundation for Growth Science in Japan (M.T.); and the Sumitomo Foundation (M.T.). We thank Dr. Terry D. Copeland for protein sequencing, Drs. Takanori Eguchi and Eiji Nakata for useful discussions, Drs. Masanao Minato and Norifumi Moritani for technical assistance, and Miss Yuki Nonami and Ayako Shizuku for secretarial assistance.

PY - 2002

Y1 - 2002

N2 - Connective tissue growth factor/hypertrophic chondrocyte-specific gene product 24 (CTGF/Hcs24) displays multiple functions in several types of mesenchymal cells, including the promotion of proliferation and differentiation of chondrocytes. Recently, the internalization and intracellular function of CTGF/Hcs24 were indicated as well. In this study, a binding protein for this factor was purified from the cytosolic fraction of human chondrosarcoma-derived chondrocytic cell line (HCS-2/8) by CTGF/Hcs24-affinity chromatography. The apparent molecular weight of the protein was 42 kDa and determination of the internal amino acid sequence revealed this protein to be β- or γ-actin. An in vitro competitive binding assay of 125I-labeled recombinant CTGF/Hcs24 with cold-rCTGF/Hcs24 showed that the binding between actin and 125I-CTGF/Hcs24 was specific. Immunoprecipitation analysis also showed that CTGF/Hcs24 bound to actin in HCS-2/8 cells. However, rCTGF/Hcs24 had no effects on the expression level of γ-actin mRNA or total actin protein. These findings suggest that a significant portion of intracellular CTGF/Hcs24 may regulate certain cell biological events in chondrocytes through the interaction with this particular cytoskeletal protein.

AB - Connective tissue growth factor/hypertrophic chondrocyte-specific gene product 24 (CTGF/Hcs24) displays multiple functions in several types of mesenchymal cells, including the promotion of proliferation and differentiation of chondrocytes. Recently, the internalization and intracellular function of CTGF/Hcs24 were indicated as well. In this study, a binding protein for this factor was purified from the cytosolic fraction of human chondrosarcoma-derived chondrocytic cell line (HCS-2/8) by CTGF/Hcs24-affinity chromatography. The apparent molecular weight of the protein was 42 kDa and determination of the internal amino acid sequence revealed this protein to be β- or γ-actin. An in vitro competitive binding assay of 125I-labeled recombinant CTGF/Hcs24 with cold-rCTGF/Hcs24 showed that the binding between actin and 125I-CTGF/Hcs24 was specific. Immunoprecipitation analysis also showed that CTGF/Hcs24 bound to actin in HCS-2/8 cells. However, rCTGF/Hcs24 had no effects on the expression level of γ-actin mRNA or total actin protein. These findings suggest that a significant portion of intracellular CTGF/Hcs24 may regulate certain cell biological events in chondrocytes through the interaction with this particular cytoskeletal protein.

KW - Actin

KW - Cell cycle

KW - Chondrocyte

KW - Connective tissue growth factor/hypertrophic chondrocyte-specific gene product

KW - Cytoskeleton

KW - Purification

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U2 - 10.1016/S0006-291X(02)02739-0

DO - 10.1016/S0006-291X(02)02739-0

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SN - 0006-291X

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JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

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ER -

CTGF/Hcs24 interacts with the cytoskeletal protein actin in chondrocytes

Abstract

Keywords

ASJC Scopus subject areas

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